Computational Biology
Over the past month, the HairDAO community has dug into computational biology and how it might help us develop new hair loss treatments. Our search for knowledge led us to explore partnerships with LabDAO, Charles River Labs, Acellera, and Insilico Medicine We were thoroughly impressed with the companies building in the computational biology space and appreciated that each company had their own angle, whether it was a proprietary model, democratized access to compute, or in-house translational capabilities
At a high-level, traditional computational biology can be broken into three sections:
1.) Target Identification or “Which gene/protein should we try to up/downregulate?”
2.) Protein Folding Prediction or “How will the protein produced by our gene be shaped?”
3.) Drug Selection or “Which drugs can bind to our target protein and have the desired effect?”
Each organization/tool plays a unique role in computational drug discovery.
Target Identification
Regarding target identification, Insilico Medicine has developed PandaOmics, an AI model capable of crawling the internet to scan relevant papers and OMICs data to suggest potential gene targets based on your indication of interest. PandaOmics then ranks gene targets based on variables like target novelty, biological relevance, confidence, commercial tractability, and safety Interestingly, as Niklas Rindtorff pointed out on Hairy Matters 4, a dedicated community of patient-researchers might actually be better suited for the target identification step in computational biology Not only are humans less prone to the hallucinations which often plague present-day large language models, but patient communities are also able to create proprietary datasets (from their own biological data), which can be used to better inform target identification and drug discovery Sorry computers, but you don’t have genomes yet!
As an example, after Jumpman suggested looking into Estrogen Receptor β (ERβ) agonists, Averbs123 suggested testing WAY-2000070, a known ERβ agonist. However, Jumpman had
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data from a group of biohackers who had already tried WAY-2000070, “They got hives but no hair”. By proceeding in that manner, the HairDAO community has generated 50 promising gene targets Even better, our patient community does not prioritize short-term profit maximization, but rather to solve hair loss alone, which we posit is the highest value accretion strategy over the long-term.
Protein Prediction
Once satisfied with our gene targets, we can either use a previously observed protein structure or tools like AlphaFold and ColabFold to predict protein shape While AlphaFold requires loads of hardware to run the required computation, ColabFold allows anybody to run its software via cloud compute. Previously, only Universities and Biotech companies (like Charles River Labs and Insilico Medicine) had access to the hardware capacity to run AlphaFold More recently, LabDAO has also democratized access to AI tooling via their PLEX product, which uses compute clusters from Bacalhau to allow users to run ColabFold from home. In the near future, LabDAO will also allow community members to provide their computational power to the PLEX network in exchange for tokens. While predictive AI like AlphaFold is very helpful to narrow the search space in computational biology, there are still limits - not the least of which is that several studies have indicated that protein shape dynamically changes in the body, and so any fixed structure presented by AI is likely to contain some margin of error.
Drug Selection
After determining a protein’s shape, AI tools like Chemistry42 have been developed to recommend drugs with high binding affinity to the target protein. While our community is also capable of recommending relevant drugs, Chemistry42’s capacity is very impressive, as it can recommend more than 8,000 small molecules when given a protein’s structure. On the other hand, PLEX allows us to test the binding affinity of various drugs using quick and dirty models like EquiBind or more accurate models like DiffDock For a best-in-class drug affinity model, Benjels got HairDAO in touch with the Acellera team, who actually won the Grand Challenge 4 Affinity Ranking Competition
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Once you’ve narrowed down your drug list to a couple small molecules, siRNA’s, or ASO’s, the next step is real world validation. For hair loss, the first validating assay would be the microdissected human hair follicle model, which ensures a drug’s delivery to the hair bulb. If successful using the microdissected hair follicle, the next step is either human scalp skin or stumptail macaque model, depending on your budget and country of residence. After that - IN VIVO, IN VIVO, IN VIVO While companies like Acellera and LabDAO outsource their validation work, we expect that Charles River Labs excels in this realm, as they supported the development of 86% of the novel drugs that were FDA-approved in 2021 and, in turn, have the most wet lab capacity of anybody in the world
The most expensive component of all HairDAO activity is our wet lab research Accordingly, our community should take on the challenge of improving their computational biology skills If we can do that, we’ll dramatically increase the probability of success each time we pull the trigger on funding a wet lab experiment Anybody who’s interested in working on computational biology should reach out to HairDAO core and we can work to get you setup on PLEX!
Update
HairDAO has engaged 5cube Labs to conduct in silico computational analysis on derivatives of a Twist-1 inhibitor Ideally, our derivative drug will exhibit increased inhibition of our target gene with an improved toxicity profile over its parent drug The in silico analysis should yield at least five derivatives, which we can then test in vitro. We’ll post all the resulting data in our Discord server behind a token-gated channel so get your HAIR soon!
Treatments
HDAC Inhibitors
Our chromosomes are made of chromatin, tightly folded packets of well-known DNA and less-rehearsed histones. Histones bind to our DNA to determine which genes are expressed in a given cell While the DNA in all our cells is the same, the histone expression of each cell differs wildly. Histones are the primary reason why some cells become brain neurons and others become hair follicles The process of cell differentiation is primarily driven by histone modifications via acetylase and deacetylase enzymes
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Histones bind to DNA to silence gene expression
Acetylation causes histones to become less attached to our DNA, thus increasing the gene’s expression that the histone previously suppressed. Deacetylation causes DNA to wrap more tightly around our histones, thus further silencing the reference gene’s expression
HATs relax our histones and consequently upregulate gene expression
Many of our researchers desire to increase the expression of genes that become silenced in hair loss, leading us to focus on histone deacetylation (HDAC) inhibition and histone acetylation (HAT) upregulation While both achieve a similar effect–increasing gene expression previously suppressed–it is significantly easier to inhibit a protein than to upregulate it–thus why there are
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a long list of HDAC inhibitors but very few HAT upregulators That doesn’t stop Metabobbly from trying HAT upregulation, which he achieves through acetyl-CoA supplementation, a co-factor necessary for healthy HAT function Metabobbly also takes 14 low-potency HDAC inhibitors In choosing HDAC inhibitors, the design space is large. There are 18 HDAC enzymes in human cells, divided into four classes.
The four HDAC classes
The most trialed HDAC inhibitor is Valproic Acid (VPA), at the top of Jumpman’s identified CMAP list of treatments and a Class I HDAC inhibitor. Similar to our T34 treatment, a low dose topical Valproic Acid treatment was applied in a small observational study in South Korea The results showed that a 65mg daily dose of topical Valproic Acid–the FDA-approved, prescribed oral dose is 500-750 mg–increased total hair count by 24 hairs per square cm versus the placebo group Jumpman also found that the FDA-approved drug lowers PAI-1 and improves the PAI/tPA ratio, both positives for hair growth.
On the other hand, Pubehead has observed a group buy test VPA without results Additionally, the side effects of VPA are severe if systemically absorbed, as HDAC alteration affects a wide range of gene targets After all, the drug was initially approved for epilepsy treatment Blackhole points out that people who take VPA orally report hair loss, but Jumpman notes it’s only a liver-activated effect We are thus left to question whether VPA regrows hair and, if it does, whether we can apply it topically without systemic absorption It may work best in combination topical therapy with Verteporfin or Minoxidil, Jumpman and Pevjo posit. Zabit Magomedov is taking it to improve his performance anxiety, while others attempt to trial it for additional medical indications
After Valproic Acid, Jumpman found that Class II HDAC inhibitors, specifically those that downregulate HDAC 4, 7 and 9, may also be worth pursuing further HDAC 4 and 9 have been found to interact with Twist1, Twist2, and Pax1, while HDAC7 interacts with the androgen receptor Our lead researcher thus hypothesizes that broad, mild Class II HDAC inhibition may pose a positive effect on our follicles, although he acknowledges their risk of side effects.
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Notable Class II HDAC inhibitors include A364365, LMK235, Sodium butyrate (a Class I and IIa HDAC inhibitor), and Vorinostat (a class I and IIa HDAC inhibitor).
Melatonin
Gorilla Head reported his initial results from his topical 6mg Melatonin treatment, “Whether it will thicken and increase density remains to be seen, but two weeks in and it’s clear it’s shutting down shedding.” As Jumpman teaches, melatonin binds to the nuclear receptor RORα, which then upregulates both B-catenin and SMAD1 B-catenin potentiates Wnt signalling, while SMAD1 upregulates RUNX2 and downregulates Twist-1 WNT upregulation, RUNX2 upregulation, and TWIST1 downregulation are several effects we seek to create in our follicles, perhaps why melatonin has appeared to separate itself from natural products that have done little for the HairForce thus far
Melatonin has other positive effects too The nutraceutical downregulates various subtypes of estrogen receptors (ER), such as ERα, which has been associated with catagen induction. Kabab added more fuel, finding that melatonin blocks Epithelial Mesenchymal Transition (EMT), whose pathways share prominent overlaps with the proteins predominant in bald scalps Visangle discovered that melatonin inhibits a wide array of SASP factors–inflammatory signals only emitted by zombie cells that can no longer proliferate but are also resistant to apoptosis–in cashmere goats
Most convincingly, melatonin has been found to meaningfully increase hair density in clinical trials by as much as 40 9% over 6 months, which seems quite high but we have no reason to distrust the referenced European doctors. Its side effect profile is relatively benign - some patients reported feeling drowsy, so it may be best to apply before bed While Gorilla Head might have overdone his 6mg dose, most studies have reported strong results with 1mg, equivalent to 1mL of a 0 1% topical solution Melatonin probably is not the cure we seek, but a low to moderate dosage may be a worthwhile addition to our stacks
Trending
Harmine: Harmine significantly reduces Twist1 protein levels in human lung and breast cancer cell lines The novel compound reduces Twist1 through protein degradation, rather than altering its gene expression. However, the treatment also inhibits DYRK1A, an enzyme required for Wnt signaling We seek to create Harmine derivatives, those which still bind to Twist-1 on its C-terminus domain but with a non-toxic off-target side effect profile.
ADQ: ADQ significantly reduces Twist1 protein levels in liver cancer cell lines Accordingly, ADQ upregulates E-cadherin and downregulates N-cadherin, the combination of which promotes cellular adhesion While Harmine is proven to inhibit Twist1 across more cell types, ADQ has shown positive results in past group buys Why did its development stop?
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Ketoconazole - Eldarlmari reports that ketoconazole upregulates PGD2 and disrupts the sebum levels of our scalp, thus causing hair loss. Jumpman counters with 30 years of human in vivo data and blames the formulation of the shampoo for Eldarlmari’s dryness Jumpman’s ketoconazole shampoo gave him great results when he applied the treatment for 30-60 minutes. VforVegeta posits its mechanism of action to be Cyp17A1 inhibition
HMI-115: Speculation surrounding Hope Medicine’s prolactin receptor (PRLR) antibody continued as we await the results of their Phase 1 clinical trial Visangle highlighted that Hope’s novel approach reduces extrapituitary PRLR, such as PRLR in the hair follicle Numerous patients have tried reducing prolactin production in the pituitary gland itself–there are several treatments available–but none have shown results Herbert posited that a prolactin or PRLR siRNA administered locally to the follicle may work, and at significantly lower costs than Hope’s monoclonal antibody treatment. However, no one’s research on PRLR can beat MoeMan the 2nd–a modern rockstar who bravely enrolled in Hope’s clinical trial, despite the negative effects that come with celebrity. We are beyond thrilled to welcome MoeMan to the HairDAO community Prolactin’s effect on the follicle has been researched and debated since 2006, highlighting the glacial pace that hair research has moved to date With MoeMan’s help, we will finally learn whether the peptidehormone’s receptors are worth targeting further.
C3G + C3A: Pevjo found that Gelucire, a frequently used delivery vehicle, can be used to enhance and maintain the stability of C3G, a close relative to our long-sought after C3A treatment Pevjo also provided a not-yet-completed task: contact T-Nation to see if they can make a C3A supplement BeansandCornbread reminded us that both C3A and C3G will eventually turn into cyanidin in the body–questioning if we should focus on C3G instead of C3A.
Cyclosporine (CsA): Cyclosporine (CsA) only grows hair in a very narrow dose range, Jumpman reported. Too much causes hair loss, too little does nothing. Herbert and Pevjo advocated for better delivery vehicles, as CsA is toxic and as such, should only be applied continuously if localized to our hair cells Jumpman pointed out that the drug upregulates both HIF-1α and PAI-1 in gum cells, which is bad if also true for our follicles. However, our lead researcher also found that the treatment significantly upregulates sonic hedgehog (Shh), the pathway Eldarmari deemed most important for hair regrowth, in our hair cells Herbert reminded us that Ralf’s work with CsA led him to focus on SFRP1 inhibition to achieve hair regrowth, although SFRP1 inhibition has yet to yield positive results in vivo Jumpman found that CsA may exert its therapeutic efficacy by inhibiting mast cell activation, perhaps the flaw in Ralf’s Wnt-based SFRP-1 approach Our lead ResearcHair shared several additional articles on topical CsA in hair and skin Should we attempt to design more CsA-based drugs in order to mimic its positive effects while reducing any potential side effects?
Tempol + CsA: Gorilla Head, TheBest, and Zero found that a Tempol + cyclosporine A (CsA) treatment may be beneficial for our follicles Tempol is a natural antioxidant, while CsA is an immunosuppressant often accompanied by toxic off-target effects. The formulations tested were 0 5% Tempol and 0 1% CsA
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Mallia Therapeutics (CD83): Follicle Thought connected us to Mallia Therapeutics, whose soluble CD83 protein treatment has shown reparative effects in murine wound healing experiments The company is now set to enter Phase 1 Clinical Trials Visangle and Jumpman found that the soluble CD83 inhibits T cell activation, and is also likely upregulated by both rapamycin and cyclosporine DNovo24 found that CD83 is upregulated in activated macrophages Despite exciting, novel mechanisms of action, Follicle Thought reported that CD83 remains several years from the clinic, for reasons unknown.
Cycloastragenol: Tigris and Eldarlmari found that Cycloastragenol upregulates Shh, FoxA2, and Wnt. Eldarmari posited a topical solution of cane sugar alcohol, water, and Cycloastragenol may penetrate our skin to reach the dermis After all, Cycloastragenol is only 490 daltons, and he can get 1g for $56 Will his self-made treatment work? Hair formulations may require a viscosity-enhancing agent, such as hydroxypropyl cellulose, for better absorption.
Halcionide: Jumpman asked why Eldarlmari doesn’t try Halcinonide instead of Cycloastragenol. Halcionide upregulates Shh by nearly as much as Smoothened Agonist (SAG) and Purmorphamine, but without constitutively activating potentially cancer-inducing SMO Turns out Eldarmari’s already tried it, but it didn’t work.
Resveratrol: Resveratrol is reported to be an inhibitor of class I, II and IV HDACs, and is widely discussed in longevity science. Metabobbly doesn’t like it though, as it’s also a Cox-1 inhibitor. Cox-1 is necessary for normal prostaglandin functioning, which is necessary for healthy follicle functioning
Finasteride: Nostro highlighted Merck’s failure to fully illustrate the effects of Finasteride Allegedly, Merck told the FDA that Finasteride’s inhibition of the 5-alpha reductase (5ar) enzyme only downregulates dihydrotestosterone (DHT) and upregulates testosterone (T), neglecting that 5ar is relevant for the production of several additional steroids, many of which are necessary for healthy brain functioning. The authors show that Finasteride also upregulates androstenedione and epietocholanaolone, while also downregulating dihydroprogesterone, dihydrodeoxycorticosterone, dehydroepiandrosterone, andorstanediol, allopregnanolone, androsterone, epiandrosterone, dihydrocorticosterone, hydrocortisone, and 11β-hydroxydihydroandrostenedione Peter Attia discussed post-finasteride syndrome (PFS) with urologist Mohit Khera. Dr. Khera teaches that Finasteride’s blockage of progesterone into allopregnanolone is the most likely cause of PFS’s brain symptoms, such as depression, anxiety, and cognition impairment Dr Khera postures that far more than 5% of Finasteride users experience side effects, but that older users conflate many of their symptoms with old age
Dutasteride: Pevjo identified which patients respond best to the highest efficacy, FDA-approved (but only for enlarged prostates) hair loss treatment If you’ve taken a deep genomics test, you can search for the single nucleotide polymorphisms (SNPs): rs72623193 in DHRS9; rs2241057 in CYP26B1; additionally, variants in ESR1, SRD5A1, CYP19A1, and RXRG are suggested to be associated with response to dutasteride
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L-ascorbic acid 2-phosphate: CatBellyRubs proposed the natural DHT inhibitor, which researchers found to reduce DHT-induced DKK1 expression in dermal papilla cells in vitro
Green Tea Polyphenols: Pevjo doesn’t have access to TM5441, but he does have Matcha Tea, which he posits may naturally inhibit systemic PAI-1 levels
Flavonoids: Herbert taught us that Flavonoids have yielded strong preclinical results on hair cells in vitro, but that they often struggle to penetrate the skin and as such, their bioavailability is too low in vivo
Metabobbly’s Stack: Metabobbly argued for more selenium, melatonin, and caffeine uptake to our follicles. He plans on maxing out on HDAC inhibitors, acetyl-coa mitochondrial nutrition, and ATP enhancement this fall. We look forward to observing his progress.
CXX5: Visangle highlighted Seoul-based CK Regeon’s treatment, a PGD2 inhibitor. Jumpman pointed out that PGD2 inhibition has been tried and failed before After all, CXX5 has only proven results in a mouse model
Igrantine-F: CaptainCalv found a potential IGF-1 upregulator that may work topically Despite Andrew Huberman’s shill of the pathway, Jumpman has shared numerous concerns about IGF-1 upregulation in the past, namely its upregulation in bald scalps.
SK-26: Jumpman identified a potentially novel PAI-1 inhibitor. The treatment significantly downregulated epithelial mesenchyme transition (EMT) signaling pathways, further emphasizing our need to bind PAI-1 before it binds to vitronectin
Gefitnib: Not only have several Gefitnib and other EGFR inhibitors been associated with trichomegaly and hypertrichosis, there are also case reports of “abundant” hair regrowth, despite their inhibition of Akt and ERK. This is considered a perplexing side effect, as Akt and ERK are positive regulators of hair growth The contradiction may be explained by NF-kB, P65, and IKK activation, also potentiated by EGFR inhibition
Lithium: Although Metabobbly explored a 50uM formulation, Jumpman warned that Lithium may cause hair loss via hair epithelial stem cell depletion.
Escin: Pevjo discovered the Wnt activator, which is posited to upregulate Fzd Would the natural product work well with a DKK1 inhibitor?
RU58841: Visangle reported that RU58841 causes more sides than Finasteride Jumpman validated why that might be the case–androgen receptor inhibition also blocks testosterone, which is actually more significant for sexual function than dihydrotestosterone (DHT)
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Tadalafil (Cialis): Pevjo and BenTD reported that If you're on an oral 5ar inhibitor, it may be smart to include low-dose Tadalafil in your protocol. They say it might reduce potential for hypoxia and fibrosis Its close cousin, Sildenafil, may be helpful for our follicles as well
Nobiletin: Jumpman and CatBellyRubs posited that the PER2 upregulator may downregulate PAI-1 and Twist-1, but hasn’t been researched much in the context of androgenetic alopecia yet
Kabab’s Natural Stack: Kabab recommended further research into several natural treatments, including Angelica Sinesis, Red Ginseng, and Caffeine
VT3989: Jumpman identified the only TEAD inhibitor that has completed a human trial The ic50 is 11nM, and the oral dose is 25 or 100mg He warns, however, that Verteporfin is a significantly safer approach in terms of anti-fibrosis therapies.
Antifungals: AubeRemix reports that topical Piroctone Olamine and Certizine (Zyrtec) may provide benefits to our follicles Both are antifungals, likely actuating a similar mechanism of action as ketoconazole
HSP27 and 90 inhibitors: Benjels and Jumpman hypothesized that Heat Shock Proteins (HSP) 27 and 90 inhibitors may downregulate Twist-1, identifying Ganetespib, Onalespib, and Luminespib as potential therapeutics. Fgsfds found that HSP27 inhibition may also significantly downregulate androgen receptor activity HSP inhibition has also been pursued to treat cancer, begging the question why more research hasn’t gone into its effects on hair cells.
AB101: Herbert identified Applied Biology’s DHT-eating bacteria treatment Visangle discovered that the drug was tested in a Phase1 clinical trial for breast cancer survivors to reduce nipple sensitivity There hasn’t been an update on their progress since 2020, leaving us with an open request for an update on the treatment’s status We shouldn’t get our hopes up, however, as the company also collaborated with Kintor Pharma on their falsified Covid treatment.
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Top 50 Targets
1 TWIST1↓: Twist1, the mother of all our gene targets, is required to initiate catagen in mice
Our research indicates that complete inhibition of Twist1 would be hugely beneficial for hair regrowth and might even be close to a cure for androgenetic alopecia. Even better, Twist1 expression has been linked to cancer metastasis, which decreases the risk of its downregulation. Twist1 is primarily activated by P65/RelA as a result of the cascade of events that occur when DHT binds to AR However, Twist1 blocks activation of NF-kB target genes, which are required to maintain hair matrix cell proliferation While Twist1 inhibition would impact nearly every gene target on our list, its primary downstream effects include upregulation of Wnt3, Fzd10, RUNX1/2/3, ERK1/2, RSPO2/3, and the downregulation of AR, TGF-β1, SERPINE1, p38/P53, FGFR1/2, and PAX1 While the effects of Twist1 downregulation would undoubtedly be good for hair regrowth, it is a transcription factor meaning it will be very difficult to drug Nonetheless, HairDAO is on the hunt to find a small molecule, siRNA, or ASO that can selectively downregulate Twist1.
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HairDAO’s Target Map, led by Jumpman
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2 SERPINE1↓: The SERPINE1 gene encodes for plasminogen activator inhibitor-1 (PAI-1) PAI-1 is a key inhibitor of fibrinolysis, the process by which blood clots are dissolved, but, at elevated levels, it can also cause cellular senescence SERPINE1’s most important role in hair loss is likely the positive feedback loop it forms with TGF-β1 and Twist1 when activated, along with integrin-mediated ERK1/2 activation In fact, the SERPINE1:TGF-β1 positive feedback loop may explain why drugs like finasteride and dutasteride do not work for some - once the feedback loop is initiated via DHT, inhibiting 5-AR upstream does not do enough to slow it down. SERPINE1 also competes with integrin β3 for binding with vitronectin, in a process that inhibits cell migration Other important downstream effects of SERPINE1 include upregulation of ICAM1 and downregulation of Wnt3a and HGF
3 EBF1↑: EBF1 directly activates Wnt10A, a Wnt ligand that is hugely beneficial for hair growth. Jumpman and Benjels identified the UK Biobank’s androgenetic alopecia dataset, which showed EBF1 as one of the disease’s most associated genes. Unfortunately, EBF1’s status as a transcription factor means that it will be difficult to drug EBF1 is upregulated by PAX5 EGFR activates mIR-221, which deactivates MAPK, which normally suppresses PAX5. As PAX5 is upregulated, the feedback loop with EBF1 increases
4. SP1↓: Specificity Protein 1 (SP1) is a transcription factor that is involved in a variety of processes, including cell growth, differentiation, and development Jumpman found that SP1 had 75 connections to other genes involved in the hair loss process, which is more than any of our other target genes. Inhibiting SP1 blocks TGF-β1 from upregulating PAI-1 in its feedback loop with SERPINE1 While not represented on our map, SP1 presence is required for prolactin to stimulate IRF-1, a bad actor itself via its role in the Twist-1 positive feedback loop. Unfortunately we have to be careful about downregulating SP1 too much, as a little bit of it is required for hair growth
5 FGF5↓: Fibroblast Growth Factor 5 (FGF5) is expressed in a variety of tissues, including the brain, heart, and skin FGF5 is a potent catagen inducer and is primarily expressed in macrophages (a type of white blood cell) found in and around the hair follicle. The short form of the protein, FGF5S, is expressed within the follicle during anagen, and antagonizes the effects of the long form, FGF5 Our lead researcher posits that induction of catagen may be due to FGF5S inhibition, which increases the activity of FGF5. While many members of the FGF family of proteins are beneficial for hair growth, FGF5 is particularly bad as it downregulates Rspo2/3, which are very important Wnt agonists. Since the short, protective form FGF5S does not contain exon2, we can target that part of the longer FGF5 protein to prolong anagen That the ORS produces catagen-inducing FGF5 supports the idea that the ORS plays an important role in regulating the cycle.
6 TWIST2 ↨: Twist2 is a member of the Twist subfamily of bHLH proteins, along with Twist1, and is responsible for the transcriptional regulation of developmental pathways in mesenchymal cell lineages (the dermal papilla and dermal sheath are both mesenchyme cells) When the Greenleaf lab at Stanford ran their integrated single-cell chromatin and transcriptomic analyses of the human scalp, they found even more linked peaks and greater frequency-matching
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probabilities for Twist2 upregulation than Twist1 related to androgenetic alopecia Shown on our map, Twist2 directly downregulates RUNX1/2 and, while it might not downregulate RUNX3 transcription, it blocks its activity While we likely want to downregulate Twist2 for the reasons above, we are less certain of it compared to Twist1, as Twist2 has biphasic effects dependent on the expression levels of Twist1/2, Id proteins, and E12/4 It has also been found upregulated in fully hairy scalps While Twist2 certainly plays an important role in androgenetic alopecia, we are not sure whether upregulation or downregulation would be better. It may depend on the time of each follicle’s cycle
7. SPI1/PU.1 ↓: SPI1 is a transcription factor gene that encodes for the PU.1 protein, which is involved in the development of immune cells, including macrophages, neutrophils, dendritic cells, T cells, and B cells PU 1 expression is found upregulated in androgenetic alopecia, downstream of PI3K and Twist1 activation. The activated PU.1 then goes on to form complexes with RUNX proteins It remains unclear what effect those complexes have on hair loss However, a recent GWAS study in June 2023 identified SPI1 as one of 25 hub genes involved in hair loss We believe that downregulating PU 1 is likely good for hair growth
8. PAX1 ↓ FOXA2 ↑: PAX1 and FOXA2 are transcription factors that are often co-expressed in tissues and cells to regulate gene expression For the purpose of hair regrowth, it is hard to tell which of the two genes is causal Eldarlmari and Tigris found that FOXA2 upregulates Sonic hedgehog (shh), Wnt5a, and Wnt7b, three prominent hair growth promoters. While FOXA2 is also involved in promoting ID proteins, it is not expressed in the dermal papilla To date, our lead researchers have hypothesized that either 1.) FOXA2 could be regulating genes far away from its own locus, 2.) the dermal papilla’s lack of FOXA2 expression is not involved in AGA, or 3.) the cell lines used in various studies are not representative of in vivo behavior In contrast, not only is PAX1 well expressed in bald scalps, but it is also upregulated by Twist1, and its activity also downregulates EBF1, which would otherwise be helpful to stimulate new hair growth Downregulated PAX1 also inhibits EGFR, which can prolong the anagen growth phase as supported by the hair regrowth achieved via pulsing EGFR inhibitors. Accordingly, we believe that we need to downregulate PAX1 or upregulate FOXA2 to stimulate hair growth Given the expression levels in the dermal papilla, and the involvement of other PAX genes in AGA, it is likely that PAX1 is the causative gene. Eldarmari and Tigriz beg to differ, however, as they test cycloastragenol, a FOXA2 upregulator, for hair regrowth
9. ICAM1↓: ICAM1, also known as CD54, is a cell surface glycoprotein that is involved in cellular adhesion - the process by which cells bind with each other to form biological structures (i.e. the hair follicle). PAI-1 (produced by SERPINE1) upregulates ICAM1 on dermal fibroblasts–the primary cell in the dermis–which allows mast cells to bind to those fibroblasts, kicking off a chain of events which leads to skin fibrogenesis (also known as microscarring, which is bad for hair growth). While ICAM1 is not a top priority target, there are available antagonists, such as Lifitegrast, which may be interesting to study further
10. RSPO2/3 ↑: RSPO2 and RSPO3 come from the R-spondin family of proteins and are transcription factors that serve as agonists of the Wnt pathway In humans, R-Spondins prevent
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the degradation of Frizzled receptors by ZNRF3 and RNF43 (known Wnt antagonists), and thus potentiate Wnt signaling. There’s no use for inhibiting Wnt inhibitors without upregulating RSPO2, as otherwise Wnt Frizzled receptors remained inaccessible to Wnt ligands– as Jumpman diagnosed Dalosirvat’s failure. While not shown in our map, RSPO2 directly upregulates Wnt2 RSPO2 is also the top autosomal hit in the Hagenaars et al 2017 GWAS analysis, supporting its significance in both male and female pattern hair loss We do have safety concerns about upregulating RSPO2/3, due to the potential of cancer formation. However, topical RSPO2 did not cause any tumor-like developments compared to Wnt3, which did cause tumors in a 2016 study on mice R-spondins also form a complex with LGR4 that prevents b-catenin degradation. While RSPO1/4 also promotes hair growth, only RSPO2/3 binds to ZNRF3/RNF43 and GPC1, making RSPO2/3 more potent Furry fact - RSPO2/3 plays a direct role in hair shaft diameter and hair length in dogs
11 ZYX↓: Zyxin (ZYX) is an actin-interacting protein essential for cell adhesion and migration Herbert found that ZYX inhibition promotes hair growth in mice and in human hair follicles in vitro Pubehead highlighted the ability for Zyxin inhibition to alter HIF-1α pathway-related genes (ALDOC/TIMP1/LDHA), furthering his and Farnsworth’s hypothesis that we should look downstream of HIF-1α signaling. Overall, downregulating ZYX corresponds with prolonging the anagen phase of hair growth Twist1 upregulates ZYX, which in turn inhibits SOX2, CD133, and NOG - all genes that are expressed by healthy stem cells Accordingly, while ZYX downregulation may be redundant downstream of TWIST1 downregulation, it could be a more druggable target, making it worthwhile for further study Pubehead and Herbert added that Zyx inhibition not only downregulates HIF-1α pathway-genes, but also Pi3k, MAPK, and TGFβ-HIPPO signaling in dermal papilla cells. ZYX expression was found to be elevated in the affected frontal hair follicles of individuals with hair loss, but not in occipital follicles (those on our sides).
12 PRKCA/PKC↓: Protein Kinase C Alpha (PRKCA) is a gene that is part of the PKC family and encodes a protein kinase that is involved in many cellular processes, including cell adhesion, transformation, and cell cycle checkpoint PRKCA is upregulated by LCK/LYN and upregulates the PI3K and ROCK pathways, ultimately upregulating Twist1 It mediates hyperphosphorylation, also known as hyperactivation, of EGFR, also bad for hair growth. Accordingly, we would like to downregulate PRKCA and there are small molecules available to do so, including Adipotide, BIM 1, AR-170, BI-6727, and more. Zooming out to other PKCs, Jumpman posits broad PKC inhibitors may be a viable treatment, due to strong regrowth he witnessed researching the protein family TCP helped verify Jumpman’s thesis and the resultant chain of events, finding that TGFβ instructs mTORC2 to activate PKCβII for increased TWIST1 expression in proximal tubular epithelial cells The Best, Zero, and TCP found that topical procyanidin B2, a broad PKC inhibitor, yields results in vivo in 60-70% of patients who tested the compound, furthering our bullishness on PKC inhibition. As Jumpman reminded us, PKCs are the most important positive regulator of Twist-1
13. PI3K↓: The PI3K pathway is a complex and highly regulated pathway, meaning there are several different factors that can affect its activity While our map shows that PI3K is primarily
20
activated by PRKCA, before PI3k goes on to upregulate IRF4/IRF1 and NF-kB, there is more to the story. Pevjo found that Pi3k is necessary for the production of IL-15, an inflammatory cytokine that Paus et al recently found prolongs anagen in human scalp follicles in vitro Dnovo found that activation of the PI3k/AKT pathway inhibits CASP3, which may also protect dermal papilla cells EGFR activation also upregulates PI3K, and Endothelin-1 likely acts through PI3K activation to trigger mast cells, sheath contraction, and ~90% PAI-1 upregulation Jumpman acknowledges that Pi3k has benefits, as it’s necessary for the proliferative effects of the VEGF pathway PI3K’s benefits mean that we would prefer to normalize its expression level rather than fully suppress the gene
14 PDGFRb/PDGFa ↑: Platelet-derived Growth Factor alpha (PDGFa) primarily binds to Platelet-derived growth factor receptor beta (PDGFRb) in the follicle There is evidence that both PDGFa and PDGFRb promote anagen and activate hair follicle stem cells. They can form heterodimers or homodimers with each other We do not know which dimerization is involved in hair loss, but GWAS analyses have indicated that both proteins play a role. PDGFRb is expressed in the mesenchyme, while PDGFa is produced in the epithelium Farnsworth countered that PDGF signaling is not necessary for hair follicle induction–but, as he and Jumpman highlighted, hair follicle induction and anagen prolongation are two disparate problems to solve Interestingly, PDGFa upregulates EGFR, which itself is necessary to initiate anagen but after consistent upregulation, inhibits follicle growth and plays a role in the early onset of catagen. While our current belief is that we would like to upregulate PDGFa and PDGFRb to stimulate hair growth, these target genes certainly warrant more study
15. IRF4/IRF1↓: Most of the Interferon Regulatory Factor (IRF) family are involved in androgenetic alopecia, with IRF4 and IRF1 being the most significant IRF4 & IRF1 can become upregulated to the point of increasing interferon response, which stimulates the immune system. In a 2023 bioinformatics analysis, IRF1/3/4/8 were all found to be among the top motifs for upregulated genes in premature hair loss However, another analysis found IRF1/2 as the top motifs in the promoters of downregulated genes. IRF2 inhibits IRF1-mediated gene transcription As we know from conditions like alopecia areata, an overstimulated immune system can be a death sentence for an otherwise healthy hair follicle The primary downstream effect of IRF4 and IRF1 is to upregulate Casp3, which is known to trigger apoptosis and is highly correlated with early onset catagen While downregulation of IRF’s could be an attractive target for hair regrowth, some IRF proteins are involved in the production of melanin and so downregulating them too much would likely cause hair graying.
16. EPHB1↓: Jumpman discovered the Ephrin (Eph) family to be one of the most enriched pathways in a miRNA analysis of a bald scalp Of the Ephrin (Eph) receptor family, Ephrin type-B Receptor 1 (EphB1) has been shown in GWAS to be associated with the androgenetic alopecia phenotype. Our lead researcher found that the Ephrin pathway is another feedback loop which can reinforce suppression of Wnt signaling in AGA The EphB1:EphB3 complex antagonizes NF-kB and Wnt signaling, and is then further upregulated by loss of Wnt activation Miniaturizing follicles may perpetually be stuck in the Ephrin-Wnt negative feedback loop. Ephrin receptor signaling is also the most enriched pathway in extracellular matrix (ECM) stiffness,
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correlating the target with Twist1 inhibition Jumpman also acknowledges Ephrin’s cross-talk to fibroblast growth factor (FGF) proteins, another prominent hair loss target group. It’s a complex target, our lead researcher warns Should we test Ephrin inhibitors further?
17 PRLR↓: Prolactin Receptor (PRLR) is a member of the cytokine receptor superfamily that is activated by prolactin (PRL), an upregulated hormone in male balding scalps Notably, PRLR--the receptor that HMI-115 inhibits–is not altered in AGA, although PRL is. Jumpman laments that we do not know the exact mechanism for PRL’s upregulation However, he posits several mechanisms: 1) the Calcineurin pathway via intracellular calcium accumulation; 2) Nerve Growth Factor (NGF) from mast cells; and 3) DHT’s increase of the binding affinity of PRL to the PRLR by altering the pH surrounding our hair cells Increased PRLR activity, driven by altered prolactin activity, renders the upregulation of LCK/LYN and downregulation of BCL6 LCK/LYN upregulation is critical in the activation of mast cells, which are bad for hair growth. However, PLRL’s downregulation of BCL6 may be good, as a significant number of downregulated genes in bald DPCs are BCL6 target genes, such as Blimp-1. In Reviewed by Ralf, Ralf warns that even topical PRLR antagonists (e g small peptide antagonists) could turn out be a shot in the foot: hair follicle epithelial stem cells actually require PRLR-mediated stimulation for survival and optimal function ex vivo; PRLR signaling also appears to be required for the expression of several functionally important hair keratins Thus, blocking or antagonizing PRLR signaling could exert undesirable long-term adverse effects even in the hair follicle itself Ralf has also shown that prolactin actually keeps female scalp hair follicles longer in anagen ex vivo, in striking contrast to male hair follicles where the opposite happens Not included on our map, Twist2 knockdown can strongly upregulate PRLR. The HairForce is still not sure how to best approach the target.
18. RUNX1/2/3 ↨?: Overall, the RUNX family of transcription factors strongly potentiate Wnt signaling, although RUNX3 can inhibit β-catenin (a promoter of Wnt signaling) RUNX proteins are necessary for cell fate specification in many tissues RUNX2 may be a regulator of sonic hedgehog signaling (Shh) in skin, as it is in bones and teeth. Twist-1 negatively regulates RUNX2 by blocking Runx2’s binding to DNA Twist-1 inhibition therefore may not only elevate RUNX2, but also Shh, what ElDarmari posits as the most important pathway for our hair’s regrowth. The expression of Runx2 mRNA was also markedly elevated in response to exogenous BMP2, reinforcing our thesis to upregulate BMP signaling Not included in our map, COX2 downregulation can also inhibit RUNX proteins. When upregulated, RUNX1/2/3 directly bind and antagonize the AR and Twist1/2. RUNX proteins are mutually antagonistic with all three of these transcription factors However, the aberrant behavior of RUNX3 makes us less confident that broad RUNX upregulation is the answer.
19 ID1/2/3/4 ↑: ID levels determine the amount of free E12 and E47 protein that is available to dimerize with Twist1, and thus can determine which Twist1 dimer (hetero or homodimer) is formed Overexpression of ID1/2/3/4 results in thicker hair shafts as Id proteins stimulate differentiation, particularly amongst hair shaft progenitor cells An inverse correlation between IDs and proliferation becomes more pronounced during repetitive rounds of depilation-induced hair cycling in mice, until eventually Id1 cKO animals failed to regrow their hair coat Relating to
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androgenetic alopecia Id proteins are primarily upregulated by BMP2/4, ALK1, and Smad1/5/8 Id proteins would be higher on our list of target genes if they were easier to drug.
20. ADRB2 ↑: ARDB2 promotes aerobic glycolysis, precocious anagen, and activates TOP1 for DNA repair, but may increase inflammation over time Overactivation may also deplete hair follicle and melanocyte stem cells It is activated by the sympathetic nerve, which wraps around the arrector pili muscle. In late AGA, the connection between sympathetic nerve and arrector pili is lost, contributing to delayed anagen entry Upregulated ADRB2 may maintain the arrector pilli’s linkage to our follicles for longer
21 MSX1↑: MSX1 has been found downregulated 3-fold in balding scalp. MSX1 is upregulated by AUTS2, causing MSX1 to promote transit-amplifying cells via upregulation of RUNX2 Not shown in our map, MSX1 also promotes BMP4, which we believe to be good for hair growth. Accordingly, we would like to upregulate MSX1 to stimulate hair growth
22 TRPS1↨: TRPS1
is a transcription
factor
that can induce excessive hair growth or hair loss
When downregulated in Ambras syndrome, TRPS1 causes hypertrichosis (excessive hair growth). However, when downregulated in trichorhinophalangeal syndrome, TRPS1 causes hair loss TRPS1 directly represses SOX9, which is necessary for stem cell specification and promotes the hair follicle fate during embryogenesis TRPS1 is a tumor suppressor gene, meaning that, even if downregulation could regrow hair, the risk may be too large. Shown on our map, TRPS1 downregulates SERPINE1 and upregulates LHX2 Not shown on our mapping, TRPS1 also binds to the promoter regions of PRDM1, SOX18, and SOX21. While an important gene in androgenetic alopecia, we are unsure whether to upregulate or downregulate TRPS1. Maintaining it at normal levels is likely the best path today
23 SFRP2 ↑?: Secreted Frizzled-related Protein 2 (SFRP2), is a biphasic modulator of Wnt activity When it binds β-catenin, but only with the C terminus, it promotes Wnt activity When it binds with the N terminus, it inhibits Wnt activity. It is also able to bind Fzd Wnt receptors to enhance Wnt signaling, or Wnt3a to inhibit Wnt activity Markedly higher SFRP2 expression is observed in beard hair follicles, followed by occipital follicles, and it was barely detected in balding frontal follicles. SFRP2 is shown to increase β-catenin accumulation in dermal papilla cells, and is correlated with trichogenicity of DPCs in vitro However, upregulated SFRP2 has been observed in premature AGA scalp DPCs. Jumpman also points out that SRFP2 is also a BMP inhibitor, which would be bad for our follicles. It remains unclear whether we should upregulate or downregulate SFRP2, although we lean towards upregulation
24 LHX2↑: LHX2 is a transcription factor that is downregulated in androgenetic alopecia LHX2 knockout has been found to cause hair follicle miniaturization and sebaceous gland hyperplasia (hyperplasia is often an early stage of cancer). LHX2 activation promotes hair follicle placode down-growth, which is the beginning of follicle neogenesis While our map shows LHX2 is upregulated by TRPS1, it is also activated by NF-kB LHX2 also acts downstream to upregulate SOX9 and Tcf4 in bulge cells, and to downregulate Lgr5 in the mesenchyme. Upregulating LHX2 is likely beneficial for hair growth
23
25. BCL2↓: B-cell lymphoma 2 (BLC2) is normally anti-apoptotic, although it has been found highly upregulated in hair loss and can promote the catagen phase of the hair follicle BCL2 is primarily upregulated by P65/RelA in our follicles. After it has been activated, Bcl-2 can form a complex with Twist1, which facilitates nuclear transport of Twist-1 and thus markedly potentiating Twist1’s hair loss effect While we likely want to downregulate BCL2, we must be careful of oversuppression, which would cause apoptosis resulting in hair loss.
26 P65/RELA↓: P65 and RelA are transcription factors which typically form a heterodimer together as part of the NF-kB pathway. NF-kB activates P65/RelA, which upregulates Serpine1, Twist1, Twist2, and BCL2–all targets we’d like to inhibit RORa can downregulate P65/RelA Notably, melatonin has been observed to downregulate P65 While we would like to downregulate P65/RelA, total knockout would probably be counterproductive, as the broader NF-kB pathway does have positive effects on hair regrowth If we can take TWIST1 out of the equation, we think P65 could actually be beneficial for our follicles.
27 AUTS2 ↑?: Activator of Transcription and Development Regulator 2 (AUTS2) is a transcription factor that is related to neurogenesis, synaptic plasticity, and neuroprotection. In hair loss, the primary mechanism of action for AUTS2 is its upregulation of MSX1, which upregulates RUNX1/2/3, which downregulates Twist1 While we feel confident that AUTS2 should be upregulated, we are not certain because AUTS2 is a known BMP repressor. However, the impact of AUTS2 on BMPs is likely minimal
28. IGFBP3/5↑: As the name suggests, Insulin-like growth factor binding proteins (IGFBPs) play a crucial role in binding IGF proteins They typically interfere with insulin signaling by reducing attachment of IGF-1 to its receptor, which will reduce mTOR activation. Additionally, IGF1, IGFBP3/5, and vitronectin form a complex that binds to integrins, regulating cell migration, which can play both positive and negative roles in hair loss However, the upregulation of SERPINE1 in AGA interferes with the IGF1:IGFBP3/5:Vitronectin complex by competing with IGFBP3/5 for binding to vitronectin, as well as interfering with binding of uPA or vitronectin to integrin avb3 While we believe that upregulating IGFBP3/5 should be good for hair growth as it is meaningfully downregulated in a balding scalp, it remains an open question which complex we should encourage and which we should inhibit–-IGF1:IGFBP3/5:Vitronectin or SERPINE1:Vitronectin?
29 PRDM1(BLIMP1)↑: PRDM1 encodes the protein Blimp1 It is a transcription factor that plays important roles in cell differentiation and development. Blimp1 deletion has been found to decrease matrix cell proliferation, cause aberrant medulla cell organization in awl3 hair follicles, and shorten the hair follicle anagen phase Per our map, Blimp1 is primarily upregulated by PI3K and it then downregulates BCL6. Not shown so clearly on our map, Blimp1 downregulation also downregulates Wnt/β-catenin signaling, which is essential for hair growth Accordingly, we likely want to upregulate Blimp1
24
30 HAAO↓: HAAO catalyzes the conversion of 3-hydroxyanthranilic acid into quinolinic acid, and is positively correlated with cellular senescence, which is terrible for hair growth. Senescent cells release SASP factors that promote inflammation, which damages hair follicles and also can compete with healthy cells for nutrients and space. Knockdown of HAAO extends lifespan by 30% and delays aging in roundworms, so the effect on senescence appears highly significant Accordingly, we would like to downregulate HAAO
31 SFRP1↓: Secreted Frizzled-related Protein 1 (SFRP1) is a known Wnt ligand inhibitor, first proposed as a potential target for treating AGA by Professor Ralf Paus Included in our map, SFRP1 can be downregulated by HGF, which is unfortunately downregulated by SERPINE1 and IQGAP1 in AGA As a result, upregulated SFRP1 suppresses both Wnt3a and Wnt10a, which are both essential for hair growth Not shown in our map, SFRP1 upregulation may inhibit Twist proteins, but only when stimulated by Wnt activation, and so is negligible as Wnt diminished in balding follicles WAY-316606 is an SFRP1 inhibitor that biohackers have tried using to treat androgenetic alopecia, but they were not successful. Nonetheless, we believe that SFRP1 downregulation is beneficial for hair growth
32. OPHN1↓: On the X chromosome, Ophoffin 1 (OPHN1) has the second strongest association with the androgenetic alopecia phenotype of any other gene besides androgen receptor (AR) expression OPHN1 upregulates AR, bad for our follicles, but downregulates the Rho pathway, perhaps good for our follicles. Shockingly, OPHN1 overexpression may actually upregulate AR even in the absence of androgens, countering the effects of drugs like finasteride and dutasteride. While not shown on our map, some of our researchers have hypothesized that Twist1 overexpression could upregulate OPHN1. Accordingly, we would like to downregulate OPHN1, but not so much that Rho becomes activated
33 RORα↑: Retinoid-related Orphan Receptor-α (RORα) is a transcription factor expressed in hair follicles that enables tissue regeneration While RORα does activate SERPINE1, which is bad for hair growth, its effects are primarily positive through its inhibition of P65, which has a multitude of positive effects for hair growth including Twist1 inhibition Further, downregulation of RORα is implicated in atherosclerosis, which is associated with AGA Accordingly, we would like to upregulate RORα.
34. ALX4↓: Aristaless-like Homeobox 4 (ALX4) is a transcription factor, which may be a downstream redundancy of Twist1 but could be easier to drug. Twist-null embryos have been found to also have ALX4 suppressed When activated, ALX4 downregulates NCAM proteins, which play an important role in cellular adhesion and signaling that is beneficial for hair growth. Not shown on our map, ALX4 has been found to downregulate the Shh pathway as well as β-catenin, both of which are good for hair growth Accordingly, we would like to downregulate ALX4.
35 GPC1↑: Growth Plate Chondrogenic Protein 1 (GPC1) is downregulated in hair loss It is an Heparan Sulfate Proteoglycan (HSPG), which regulates stem cell fate and differentiation. It binds to RSPO2/3 to downregulate ZNRF3/RNF43, which would otherwise degrade Fzd, a
25
critical Wnt receptor Not shown on our map, GPC1 also mediates VEGF/HGF-induced angiogenesis, which is good for hair growth. Accordingly, we would like to upregulate GPC1.
36. DKK2↨: Dickkopf-related Protein 2 (DKK2) is another protein with biphasic effects on Wnt signaling dependent on context It binds to co-receptors LRP5/6 to inhibit Wnt signalling, a definitive negative However, it either potentiates or antagonizes the effects of DKK1, which can be good or bad for Wnt. DKK2 knockout mice can regrow hair, despite SOSTDC1 activation partially restricting growth Downregulation of both DKK2 and SOTDC1 could strongly promote hair growth, our lead researcher posits We lean towards downregulation of the target, although we remain uncertain due to DKK2’s downregulation in bald scalps.
37 DKK1↓: Dickkopf-related Protein 1 (DKK1) is a protein that binds to the Wnt receptors LRP5/6, which, like DKK2, inhibits the ability of Wnt3a and Wnt10a to potentiate Wnt signalling. However, inhibiting the protein alone doesn’t do much for us We need to pair a DKK1 inhibitor with a Fzd receptor upregulator, as more LRP5 is useless without more Fzd. Interestingly, Surrozen has actually developed a modified LRP/Fzd antibody using modified DKK1 to mimic Wnt3 upregulation, as Wnt3 itself insoluble
38 ANGPTL7↑: Angiopoietin-like Protein 7 (ANGPTL7) is a protein that has been shown to regulate angiogenesis and lymphatic drainage in our hair bulge stem cells, both good for healthy follicle functioning. ANGPTL7 blocks the inactivation of FOXO1, which we also want to upregulate, and has also been found highly downregulated in AGA GWAS Shown on our map, SERPINE1, which we want to downregulate, strongly downregulates ANGPTL7. Not shown on our map, Id1 knockdown has been found to suppress ANGPTL7. Additionally, ANGPTL7 activation has been found to upregulate AXIN1/36/37, TCF3/4/41, LEF1/39, CCND1/40, and MYC42–all of which help our follicles grow. We want to upregulate ANGPTL7.
39 SOSTDC1↨: Sclerostin Domain Containing 1 (SOSTDC1) enhances insulin secretion and glucose homeostasis. One study found that it is only expressed in bald dermal papilla cells. Shown on our map, SOSTDC1 activation downregulates BMP2/4, ALK1, SMAD1/5/8, and Id1/2/3/4 expression, which activates Twist1 and is bad for our follicles Not shown on our map, SOSTDC1 is also known to inhibit LRP5/6, reducing Wnt activation. We likely want to downregulate the target, although we should be weary of how inhibiting SOSTDC1 effects SOX2 activity.
40 CASP3 ↓ or DFFA ↓: Caspase-3 (CASP3) and DNA Fragmentation Factor Subunit Alpha (DFFA) are both proteins that play a meaningful role in the process of apoptosis, and their expression is highly correlated to androgenetic alopecia CASP3 is activated by a number of factors, including stress, inflammation, and DNA damage DFFA is a substrate of CASP3 that is associated with AGA in GWAS. It is activated by CASP3-mediated cleavage, and then fragments DNA during apoptosis Shown on our map, CASP3 is primarily downregulated by HGF and upregulated by IRF4/IRF1 Not shown on our map, VEGF protects hair bulge stem cells, inhibiting PI3K, preventing it from upregulating CASP3 along with Bax, Twist1, and Serpine1 We would like to downregulate CASP3 to stimulate hair growth
26
41. EPS8↓: A substrate of the epidermal growth factor (EGFR) pathway, Epidermal Growth Factor Receptor Pathway Substrate 8 (EPS8) is typically bad for hair growth Shown on our map, EPS8 strengthens EGFR signaling (a known catagen-inducer), and EPS8 serves as an essential protein to activate the RAC pathway, which upregulates Serpine1 EPS8 is also upregulated by P65/RelA Interestingly, estrogen downregulates EPS8 by more than half in male hair follicles, while it has no inhibitory effects on EPS8 in female hair follicles. Finally, EPS8 knockdown demonstrated upregulation of E-cadherin, downregulation of N-cadherin, and vimentin - all positive effects for hair regrowth Accordingly, downregulating EPS8 would be beneficial for hair growth.
42 HDACs particularly type II 4/9 ↓: Histone Deacetylases (HDACs) are proteins that remove acetyl groups from histones, which would otherwise help package DNA into chromosomes. HDACs will typically suppress their target genes While not fully represented on our map, we believe that HDAC4/9 likely suppress the gene expression of an AR inhibitor, resulting in AR upregulation However, in prostate cancer, HDAC4 does bind directly to AR, resulting in AR inhibition HDAC4/9 are also recruited to DNA by Twist1/2 to downregulate gene transcription
Broad HDAC inhibitors, such Vorinostat, have been tested in vivo unsuccessfully in attempts to treat androgenetic alopecia However, that may speak more to the binding affinity of the drug than to the efficacy of downregulating HDAC4/9 II to regrow hair
43 BCL6↑: B-Cell Lymphoma 6 (BCL6) is a transcription factor that is commonly found downregulated in androgenetic alopecia. BCL6 promotes survival of hair cells, and specifically promotes the cell proliferation and differentiation that is involved in the growth of the hair shaft. Prolactin strongly suppresses BCL6 Downstream, BCL6 activation downregulates Blimp1, but Blimp1 activation also downregulates BCL6. While Blimp1 activation cooperates with Wnt signaling, suggesting a negative byproduct of BCL6 activation, we believe that upregulating BCL6 will likely do more good than bad for hair growth As tangential evidence, downregulation of BCL6 results in the degradation of cochlear hairs in the ear, which are essential for hearing. However, BCL6 upregulation is also associated with cancer, and so we likely would only want to normalize the protein from its downregulated levels
44 15-PGDH↓: 15-Hydroxyprostaglandin Dehydrogenase (15-PGDH) is an enzyme involved in the metabolism of prostaglandins. As shown on our map, 15-PGDH activation downregulates RUNX1/2/3, which would otherwise play an important role in downregulating Twist1 and AR. Not shown on our map, 15-PDGH inhibition increases tissue PGE2 levels, which aids tissue regeneration. As such, 15-PGDH inhibition increases Type IV collagen, improves re-epithelization, and reduces scarring in wound healing However, 15-PGDH knockout mice have a 7 6-fold increase in colon tumors, which gives us pause to pursue 15-PGDH inhibition Amgen does have a 15-PGDH small molecule inhibitor, SW033291, in clinical trials to regenerate colon, liver, and bone marrow tissue While it appears unlikely that Amgen will put the drug through clinical trials for AGA, it is patented for hair loss
27
45 ALK5↓: Activin Receptor-like Kinase 5 (ALK5) is a receptor to TGF-β1 signaling Once activated by TGF-β1, ALK5 then activates Smad2/3, which downregulates Smad1/5/8 and Id1/2/3/4 while activating Sp1 and P21 Small molecule TPO427736 inhibits TGF-β1 downstream of androgens via Alk5 suppression. However, Alk5 does play a role in angiogenesis, necessary for healthy follicle functioning, rendering that moderate downregulation of Alk5 is likely more beneficial to treat androgenetic alopecia than full inhibition
46 YAP1 or TEAD1/2/3/4 ↓?: Yes-associated Protein 1 (YAP1) is a transcription factor that activates TEAD1/2/3/4 transcription factors and has a significant effect on wound healing In hair transplants, injections of renowned YAP-1 inhibitor Verteporfin may decrease fibrosis and increase the odds of follicle neogenesis at the extraction site Aspect reminds us fibrosis may be the only way to never regrow our follicles, ratified by Dr Ralf Paus and Dr Maksim Plikus long ago, with Jumpman hypothesizing that we need to explore topical YAP1 inhibitors further. Farnsworth agreed, finding that increased extracellular matrix (ECM) stiffness elevates YAP/TAZ nuclear localization, and that TGFβ2 increased nuclear YAP/TAZ in both normal and glaucomatous HTM cells However, YAP1 inhibition is correlated with cyst formation, making our path forward not entirely clear While our map shows that YAP1 activation upregulates SERPINE1, YAP1 is involved in several interactions not shown. Focal adhesion kinase protein (FAK) and CASP3 upregulate YAP1, which then downregulates COX2 and blocks ERα transcription of ERS1, both good and bad for hair growth We need more research into YAP1 and TEAD proteins in both wounding and non-wounding environments.
47. Notum↓: Notum inhibits Wnt signaling through the downregulation of Wnt3a and Wnt10 ligands. Downregulating Notum prevents the degradation of Wnt ligands, but unfortunately the impact of Notum downregulation on hair loss would be muted if the Wnt Fzd receptors are still blocked, a similar problem encountered with DKK1 inhibition. Similarly, while Notum may participate in the initiation of catagen, it is not required to initiate catagen Nonetheless, moderately downregulating Notum likely benefits hair growth
48 LCK+LYN↓: Tyrosine Kinases LCK and LYN promote inflammation, macrophage infiltration, and activation of mast cells Shown on our map, LCK and LYN are primarily upregulated by TGF-β1, PRLR, and the Rho pathway. Activated LCK/LYN then also upregulates Rho, creating a positive feedback loop Activated LCK/LYN also upregulates PRKCA and Smad2/3, both bad for hair growth. Given the known downstream effects of LCK/LYN activation, downregulation is likely beneficial for hair growth.
49. NR3C1↑: Nuclear Receptor Subfamily 3 Group C Member 1 (NR3C1), also known as the glucocorticoid receptor, encodes a protein that binds to the stress hormone cortisol and decreases its effect When downregulated, NR3C1 can cause hirsutism and alopecia in women
Despite that NR3C1 can upregulate SERPINE1, which is bad for hair growth, we believe that upregulating NR3C1 would do more good than bad in terms of stimulating healthy hair growth It was found downregulated in bald scalp, and one analysis found it to be the number two hub gene in AGA.
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50 SPAG17 ↑?: Sperm Associated Antigen 17 (SPAG17) plays a role in the assembly and functioning of cilia, hair-like structures that project from the surface of cells. SPAG17 had the single strongest SNP probability in a single cell chromatin analysis conducted on various hair follicle cells. Interestingly, while SPAG17 is found upregulated in the dermal papilla, it is downregulated in the dermal sheath, where Dr Kevin McElwee thinks we should research further Genetic ablation of SPAG17 has been found to promote fibrosis and extracellular matrix, hindering our follicles and further strengthening our belief that we want to upregulate the target.
More Targets
EGFR: Thanks in large part to Farnsworth, our discussion turned towards the role Epidermal Growth Factor Receptor (EGFR) plays in hair loss. EGFR is a protein found on the surface of cells that typically binds with its ligand, epidermal growth factor (EGF) While EGF doesn’t bind to the Androgen Receptor (AR), EGF has been found to inhibit AR transcription As EGF is downregulated in AGA, there appears to be a cross-talk malfunction between AR and the EGF pathway, which allows for repressed EGF production but unrepressed EGFR activation in our follicles. Of concern, the same cross-talk malfunction occurs between EGFR and AR in prostate cancer cells
Both in vivo and in vitro data support EGFR inhibition. In vivo, there have been multiple reports of hypertrichosis induced by EGFR inhibitors Jumpman reported increased hair regrowth while experimenting with Erlotinib pulse therapy Yoshimura and Ejima also reported two cases of hypertrichosis in patients taking Gefitinib, another potent EGFR inhibitor used to treat certain cancers In agreement with our in vivo findings, Jumpman found that EGFR suppresses Wnt/β-catenin signaling, and that EGFR activation may also be required for the proper function of the TGFβ1 → PAI-1 feedforward loop, both of which cause hair loss. Finally, EGFR activates mIR-221, deactivating MAPK, which activates PAX5, forming a positive feedback loop to increase EBF1 expression. EBF1 is yet another gene at the top of our target list that we would like to downregulate, further strengthening the argument for EGFR inhibition
However, there are also strong arguments in favor of EGFR upregulation. Farnsworth shared that EGF and EGFR activation in the outer root sheath (ORS) prevented the onset of catagen in mice Jumpman found that TWIST1 inhibition downregulates PAX1, which upregulates EGFR While there was some debate around EGFR’s relationship to Nuclear Factor Kappa Beta (NF-kB), we settled on the consensus that EGFR likely activates NF-kB (except in some oncogenes) which has both positive (miR186 upregulation) and negative (p65 upregulation) effects on hair loss The gene target sits on what we would call the frontiers of hair loss research–in a land where there are still more questions than answers Accordingly, we posit that EGFR is well worth further research, as theory will only take us so far.
Shh: Sonic hedgehog (Shh) signaling is commonly thought to improve our hair growth ThomasB and Visangle hypothesized that Technoderma’s TDM105-795 may work by promoting Wnt and Shh Also in trending hair loss research, Herbert and Jumpman highlighted Amplifica’s recombinant SCUBE3 treatment, a protein whose bullish thesis is predicated on its upregulation
29
by Shh Herbert shared a new paper on Shh in hair bulge stem cell culture, showing that increased Shh signaling significantly upregulates their proliferation. However, significant debate ensued between David Enshel, Eldarmari, and Jumpman on just how important Shh really is–from our hair’s most important pathway to a fairly irrelevant co-factor.
David Enshell found in mice that at telogen, Shh transcripts are absent During the transition from telogen to anagen, Shh expression is induced in the hair germ’s newly formed transient amplifying cells (TACs) Shh from the TACs is required for bulge stem cell activation, necessary to allow the bulge to differentiate into the outer root sheath, the future bulge, and the next hair germ. Due to its importance in epithelial stem cell activation, Enshell finds that ablation of Shh in the epithelial part of the hair follicle during telogen resulted in a complete lack of hair follicle regeneration
However, while stem cell proliferation within the bulge is diminished, a burst of initial proliferation within the germ still occurs in the absence of Shh. Enshell teaches that the initial anagen induction is consistent with the dermal papilla as a source for the signals that induce proliferation within the hair germ, suggesting that Shh is dispensable for anagen induction by the dermal papilla (DP). The DP and Matrix respond to Shh differently–in fact, Enshell finds that too much Shh causes non-canonical Wnt activity in the DP, causing hair loss
Eldarmari added that Shh is the most downregulated pathway in hair loss. He hypothesized that Shh upregulates Wnt, citing his own chain of events FoxA2 = Shh + Wnt5a +Wnt7b = Gli2 + Sox9 + Frizzled 4 = R-spondin = B-catenin = Wnt. In agreement, Paus et. al list Shh as an initiator of anagen in the inner root sheath and dermal papilla, venturing one step further than Enshell
Jumpman counters, however, that GLI1, a primary Shh target gene, has not been found downregulated in hair loss Jumpman reminds us that 1000s of targets upregulate Wnt, siding with Enshell that Wnt in the dermal papilla upregulates Shh in the hair matrix, which does help further potentiate Wnt signaling but is not the key driver Jumpman posits Eldarmari’s study shows that lncRNAs are not the driving force in hair loss Our current theory remains that Shh is crucial for the activation of the bulge, and thus is necessary for hair regrowth, but may negatively impact our follicles if over-activated in our dermal papilla, through the potentiation of non-canonical Wnt signaling.
miR-221: Farnsworth and dNovo24 found that androgen receptor activity promotes miR-221, which then suppresses IGF1. IGF1 has a tentative history in our research, and so our miR-221 research remains open-ended However, Farnsworth also found that the miRNA downregulates TRPS1 and ERa, two proteins we seek to upregulate
ING1 and ING2: Jumpman finds that ING1 inhibits Twist-1, so we likely want to ensure its levels remain elevated Vorinostat inhibits its closely-related ING2 Vorinostat promotes cell motility, whose role in our follicles remains undefined. Accordingly, our consensus on ING2 remains open-ended
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NPAS2: Jumpman finds that downregulating the circadian clock gene promotes surgical skin wound healing in mice The NPAS2 inhibitor used is Dwn1
Gsk3b: Dnovo and Follicle Thought highlight a new hair loss drug discovered through in silico drug design, a Gsk3b inhibitor termed LCH34 Jumpman laments that GSK3b inhibition is over-done in the hair loss community, as CHIR99021 already exists and any additional improvements are likely marginal Visangle asks what the difference is between GSk3a and GSK3b–the two GSK isoforms Jumpman answers: they both phosphorylate B-catenin, but GSK3B is the predominant one in our follicles.
c-Myc: A transcription factor whose overexpression is associated with various forms of cancer, Jumpman predicts that its transcription is overactive in androgenic alopecia as well. However, c-Myc is also found to be associated with upregulated Wnt levels, and is upregulated by PDGF signaling, another alleged pro-hair pathway, leaving room for doubt. Our current hypothesis remains largely open, leaning towards the theory that we want to normalize c-Myc expression While upregulation of c-Myc is associated with androgenic alopecia, downregulation of c-Myc is associated with senescent alopecia.
RAS: Jumpman found that RASA2, an inhibitor of RAS signaling, is downregulated by DHT Pevjo also found that RAS downregulation promotes reproductive organ health. However, RAS also upregulates c-Myc, a protein we likely want to normalize We likely want to downregulate RAS, but need to ensure its downregulation doesn’t carry adverse effects.
ERK: Kinases are enzymes that transport phosphate molecules There are strong cases to downregulate Extracellular signal-Related Kinase (ERK). ERK activation upregulates expression of Twist-1, along with Pi3k/AKT activation, both of which cause hair loss It’s also found on ZeroGPT Osteopontin upregulates ERK and Twist-1 as a result, another flaw in the trending treatment. However, there are also prominent cases against downregulation. Farnsworth found that DHT inhibits the protein Relating to our prior EGFR work, EGFR inhibitors induce hypertrichosis despite inhibiting Akt and ERK How we wish to regulate the target remains open
GPR30 or GPER1: While estrogen receptors (ER) ERα and ERβ are well-studied, a third and more recently discovered estrogen receptor, the G Protein-coupled Estrogen Receptor 1 (GPER1), has been largely neglected. GPER1 has been found responsible for rapid non-genomic estrogen effects in the reproductive, nervous, endocrine, immune, and cardiovascular systems. There remains scant data on GPER1 in hair loss research. With its known interaction with estradiol–a known potent hair growth hormone–Jumpman wonders why we haven’t looked more closely at the target Perhaps even in conjunction with 17-b-estradiol and an ERb antagonist, to find which non-hormonal effects of 17-b-estradiol might regrow our follicles
CXCL12: Follicle Thought informs us that Epibiotech is developing an siRNA for CXCL12. Tigris points out that inhibiting CXCL12 should also inhibit PAI-1, one of our top targets Jumpman
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posits that both CXCL12 and CXC4, CXLC12’s downstream target in its hair loss-associated pathway, could be inhibited to our benefit.
Rho/ROCK: There are strong cases to downregulate Rho-A-associated kinase (ROCK). In the Jumpman Chain of Events, ROCK helps upregulate Pi3k, a prominent role in our lead researcher’s reaction sequence Jumpman also found that ID1 and ID3, two proteins we likely want to upregulate, may inhibit ROCK. Additionally, Twist-1 may be upregulated independent of PKC through ROCK independent activation of Smad proteins, further reason for ROCK’s downregulation Jumpman’s belief in inhibiting target led him to resist over-inhibiting OPHN1 Pevjo also found that ROCK inhibitors may help treat erectdile disfunction. Should we look further into ROCK inhibitors as viable treatments?
PER2: Jumpman finds that Twist-1 inhibits CLOCK mediated activation of PERIOD (PER) genes, as our top target suppresses the same E-box region of our DNA that activates our PER The downregulation of PER2 also upregulates PAI-1, further perpetuating the Jumpman Chain of Events Through PER2, Twist-1 may provide a link between the immune system and the circadian timing system Interestingly, Ralf Paus discovered that BMAL1, CLOCK1 and PER1 downregulation prolonged anagen, providing an interesting contradiction among the PER genes
Snail: Visangle found that co-expression of HIF-1α, Twist, and Snail in primary tumors of patients with head and neck cancers correlated with metastasis and the worst prognosis Do we seek to downregulate all three?
FGF18: The protein is often referred to as a quiescent marker by traditional hair loss science, but the HairForce–led by Visangle and his AI-assisted research tools–found evidence to both inhibit and activate FGF18 On the case for inhibition, FGF18 activates FGFR3 in hair bulge stem cells, and consequently the Mapk/ERK and Pi3k/Akt pathways downstream of FRGR3, both of which cause hair loss. On the case for upregulation, FGF18 activates SP1, EGR1, and ETS, all of which inhibit Twist1 transcription FGF18 also upregulates ID2, which binds to E12 and thus further blocks Twist-E heterodimers Due to its biphasic effects, our target level of FGF18 remains unknown.
NOX: NAPDH Oxidases (NOX) enzymes, attached to the extracellular side of our cell membranes, come in seven isoforms: NOX1-5 and DUOX1-2. Dnovo24 found that NOX4 knockout reduced ROS generation and attenuated hypoxia-induced secretion of growth factors in human embryonic mesenchymal stem cells. Our hair cells, specifically those in the epidermis, are one of the few hypoxic environments in our bodies–meaning that they are frequently exposed to low levels of oxygen Farnsworth points out that one of the only similar cell groups is our pancreatic cells. NOX inhibition has also been observed as a potential Parkinson’s treatment Do we want to downregulate at least NOX4 in our epithelial hair cells? Herbert still wants to see the NOX - DHT pathway validated in human hair follicles in vivo He might find an answer soon–APX311 is a NOX inhibitor Jumpman is currently evaluating.
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lncRNA AC010789 1: Visangle found that the lncRNA AC010789 1 is significantly downregulated in scalp tissues of patients with androgenetic alopecia compared to adjacent normal tissues He found that the long non-coding RNA upregulates Wnt, downregulates TGFb, and downregulates miR-21-5p. How should the we seek to upregulate lncRNA AC010789.1?
αSMA: Catagen is triggered when our dermal papilla niche is pulled upwards by our dermal sheath, which is triggered by αSMA. Catagen induces both dermal sheath contraction and epithelial progenitor cell death, such that the dermal papilla can reach the stem cell reservoir in the bulge and matrix progenitors can be refreshed for the next hair cycle Shiseido revealed last year that the response to treatment with their dermal sheath cell injections inversely correlated with α-SMA expression Twist1 and PAI-1 knockdown inhibits αSMA Should we find more ways to inhibit αSMA in our mesenchymal hair cells?
ET-1: Endothelin-1 (ET-1) also triggers dermal sheath contraction ET-1 is a mast cell activator, and it’s been shown to upregulate PAI-1 by about 90%, perhaps through the PI3K P85 subunit. Jumpman posits that if αSMA is constitutively expressed along with ET-1-mediated PAI-1 upregulation, then follicle downgrowth would be inhibited, and our hair cells would not receive their needed HGF from neighboring adipocytes. HGF, among other actions, blocks Wnt inhibitors, and so our hair cells’ inability to receive it may downregulate Wnt and cause our follicles to fail to re-enter anagen
PGD2: PGD2 is predominantly released by mast cells, which are overactive in hair loss and secrete many factors to induce catagen. Jumpman teaches that, while PGD2 is upregulated in bald scaps, polymorphisms of its primary receptor DP2’s show no correlation with hair loss. Given the lack of polymorphism association, and zero effect from 2g of daily Setipiprant (a PGD2 inhibitor), Jumpman hypothesizes that PGD2 is an insignificant contributor to our follicle’s health
NF-kB: Jumpman discovered the dichotomy of the transcription factor Nuclear Factor kappa Beta (NF-kB) NF-kB has been found essential for Shh and cyclin D1 expression NF-kB also activates P65, which then activates Twist-1 directly but also suppresses Twist-1 indirectly through miR186 activation. Twist proteins generally repress NF-kB mediated gene transcription, and exert particularly potent repressive effects in NF-kB deficient systems Could miR186’s inability to repress Twist then be at play, Jumpman wonders. If miR186 is unable to inhibit Twist, Twist may inhibit NF-kb until it is depleted, at which point Twist can run even wilder in our follicles
miR186: Jumpman illustrated that TWIST1 represses miR186 by recruiting DNMT3a and DNMT3b to hypermethylate (adding a methyl group to) the miR186 promoter Twist-1's repression of miR186 then forms a feedback loop where Twist-1 is consistently upregulated, as miR186 would otherwise repress Twist-1
P65: P65 may be a tempting target to downregulate TWIST1, as it is required for TWIST1 induction However, Jumpman advises against targeting P65, due to its knockdown of Twist-2
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Knocking out both TWIST isoforms could lead to senescence and hair loss Additionally, NF-kB, which is promoted by P65 through miR186, is important for hair growth and Wnt activity. Considering all of this, targeting TWIST1 directly is more desirable than targeting P65
EDAR and EDA2R: EDAR is found in differentiating hair matrix cells responsible for building the hair shaft The EDAR variant 370A was found to enhance NF-kB activity via EDA2R upregulation. Jumpman wonders if the EDAR 370a variant overcomes TWIST1 suppression of NF-kB more so than other variants, and thus keeps miR186 activity high enough to keep TWIST1 suppressed? The 370A variant originated in East Asia under positive selection It is found in all Native Americans, who express AGA far less than other populations. It is also found in high frequencies in East Asia, another region with strong hair phenotypes EDA2R has been determined to be associated with hair loss, with a p-value comparable to the androgen receptor How should we research EDAR further?
GATA6: GATA6 is required for EDAR and constitutive activation of NF-kB in hair matrix cells.
GATA6 knockout mice undergo G2 cell cycle arrest, increased apoptosis, and premature catagen Knockout of GATA6 reduces P65 in the hair matrix during anagen, resulting in defective early progenitor cell activation and self-renewal of mature progenitor cells. We certainly want GATA6 normalized, if not upregulated
CAGs: Gorilla Head finds that our CAG repeats get shorter as we age and our testosterone levels decrease He posits that hair loss treatments may also function as longevity treatments
The Cadherins: TWIST1 causes a loss of E-cadherin and increased N-cadherin. E-cadherin is necessary for maintaining the adhesive properties of our hair stem cell niches, as well as proper skin and hair stem cell differentiation. N-cadherin is a crucial protein in epithelial mesenchyme transition (EMT), a pathway with high correlation to hair loss Interestingly, P-cadherin made it to ZeroGPT’s target list
FOXC1: Jumpman finds that FOXC1 upregulates SFRP1 Our leading traditional researchers pose an interesting dichotomy: Ralf has taught us we want to inhibit SFRP1, but Elain Fuchs states we want to upregulate FOXC1 in the bulge, which would upregulate SFRP1. Jumpman plays arbitrator, finding that SFRP1 and FOXC1 are upregulated several fold in bald dermal papilla, signaling that we want to inhibit both, but only in our follicle’s mesenchyme compartment. FOXC1 appears to be regulated by the EMT pathway, signaling that Twist-1 inhibition may downregulate the target
SDR16C5 and SDR16C6: Jumpman finds that mice lacking the epidermal retinol dehydrogenases display accelerated hair growth Both are upregulated by tretinoin, signaling we may want to find compounds whose effects are opposite to the frequently ill-prescribed retinoids
Scd1: Jumpman finds that downregulation of Scd1 in hair matrix cells causes hair loss. Scd1 is required for autocrine Wnt signaling in the matrix, as well as paracrine Shh signaling from the
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matrix to the dermal papilla, which further potentiates Wnt in the dermal papilla Without it, we lose one of our most important pathways.
NFATC1: NFATC1 inhibition is Cyclosporine-A’s proposed mechanism of action for hair regrowth. Should we try to find more non-toxic NFATC1 inhibitors?
NOTCH: DNovo24 teaches that a fully functional Notch receptor binds to a ligand (Dll1-4 or Jag1-2) presented by a neighboring cell, resulting in a conformational change of the Notch receptor that expose the recognition site for cleavage by ADAM proteases and γ-secretase, leading to the release of the active Notch intracellular domain (NICD). Subsequently, NICD is translocated to the nucleus, leading to the formation of a complex with DNA-binding protein CSL, which induces the transcription of target genes Without the activation of Notch, the NICD does not enter the nucleus, and CSL might instead interact with ubiquitous corepressors (Co-R) and histone deacetylases (HDACs) to repress target gene transcription Consequently, Notch signaling plays a vital role in the activation, proliferation, deafferentation of HFSCs and metabolite generation
HIF-1α - VEGF Pathway: There are strong cases to both upregulate and downregulate the HIF-1α - VEGF pathway, leading us to currently believe that the pathway should be normalized On the case for upregulation, Minoxidil potentiates the Hif-1α-VEGF pathway pathway in our hair cells in a dish, a proposed method of action for one of our two FDA-approved drugs. The study showed the same effect on murine skin in vivo On the case for downregulation, the Minoxidil study has not been verified in human vivo. In fact, HIF-1α and VEGF have also been shown to be upregulated in male bald scalps. Additionally, diazoxide, a less safe but still another hypertrichosis-inducing drug, has not shown any effect on HIF-1α or VEGF Stemoxydine, an HIF-1α upregulator, does not seem to promote significant hair growth. Visangle hammers that HIF-1α promotes Twist-1, the most important target in the Jumpman Chain of Events Epitomizing the dichotomy, Cyclosporin-A, a toxic but hypertrichosis-inducing drug, was shown to upregulate HIF-1α and consequently PAI-1. HIF-1α regulates a wide variety of profibrogenic genes, which are closely associated with tissue fibrosis, along with a variety of growth factors, such as VEGF To move forward, Farnsworth assumes that we need to find ways to grow our follicles despite presence of HIF-1α, as the bulge will remain hypoxic. Pubehead signals in the same direction, arguing we should look further at how upregulated HIF-1α signaling from epithelial bulge cells impacts mesenchymal dermal papilla cells.
PHD2/EGLN1 - HIF-1α Pathway: HIF-1α subunits are tightly regulated by HIF prolyl hydroxylases (PHDs), also called EGLNs. Pubehead found that EGLN1 downregulates HIF-1α under both normoxia and hypoxia Conversely, silencing of EGLN1 expression augmented HIF-1α transcriptional activity and its target gene expression in hypoxia Pubehead also found that EGLN1 and EGLN3 are both upregulated in bald scalps. If EGLN1 upregulation downregulated HIF-1α, how could both be upregulated in our balding heads? Farnsworth clarifies that the negative feedback loop doesn’t work if pyruvate is shuttled into the mitochondria, a positive argument for Dr. Bill Lowry’s research.
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Wnt - NF-kB pathway: Wnt activates NF-kB through EDAR upregulation However, in the absence of NF-kB, Wnt activity cannot be maintained in adult hair follicles.
Trending Topics
Jumpman on Mast cells: Jumpman explains that mast cells are two-fold higher in balding scalps, with inhibition of mast cell degranulation prolonging the anagen phase of the hair cycle Release of hair growth modifying factors by mast cells is under the control of secretagogues including NGF, Substance P (promotes immune privilege collapse), and ACTH, all of which promote PAI-1 In fact, PAI-1 was recently revealed to mediate mast cell degranulation in fibroblasts via upregulation of ICAM1, and it also sharply increases α-SMA expression in dermal fibroblasts Mast cells can even attenuate the endothelin-1 (ET-1) itch–with their presence as why some people don't get the infamous DHT itch Jumpman also posits that inhibition of mast cell activation may be Minoxidil’s and Cyclosporine-A’s mechanism of action. We want to run further tests on mast cells, but Herbert laments that for some targets, such as mast cells, our in vitro models might not be as suitable, as our immune response may be significantly modulated without surrounding tissues. How do we run more mast cell tests in vitro?
On the ORS vs the DP: To Jumpman’s response that FoxA2–the target of Eldarmari’s Cycloastragenol–isn’t expressed in the dermal papilla (DP), Eldarlmari states that hair loss may have to do with the outer root sheath (ORS) more than the DP If the DP were the culprit, bald follicles would have no follicle at all, instead of the miniaturized vellus hair that remains. Jumpman counters with that the DP is the command center that controls the ORS, and that hair loss must begin in the DP because that's where the androgen receptors are In an attempt to settle the debate, David Enshell beautifully describes the hair follicle cell cycling process, which indeed manifests as a circle where it is hard to identify the chicken and the egg Paus and Stenn illustrate in their oft-cited 2001 paper that the first signal for hair follicle formation, also known as morphogenesis, is generally held to come from the dermal papilla mesenchyme. However, what initiates anagen from telogen in a mature follicle is still debated The signal could arise in the resting papilla, the resting epithelial germ, the adjacent epidermis, or, in theory, even the supportive vessels, nerves, lymphatics, and resident dermal hematopoietic cells.
miRNA Analysis: MicroRNAs (miRNAs) are a class of short non-coding RNAs that regulate gene expression post-transcriptionally Typically, miRNAs bind to the 3'-UTR (untranslated region) of their target mRNAs, resulting in the repression of protein production through mRNA destabilization and translational silencing. Farnsworth shares a list of the miRNAs most differentially expressed in hair loss Jumpman outlines the pathways most associated with Farnsworth’s miRNAs Ephrin, ErbB, and Insulin Receptor signaling are the three most implicated, just ahead of androgen signaling. That Ephrin hair research has stagnated for over a decade leads us to believe it may be worth pursuing further
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Hair Graying: Jumpman dissects hair graying. The phenotype results from activation of the sympathetic nerves that innervate the melanocyte stem cell niche Under conditions of stress, the activation of these sympathetic nerves leads to burst release of the neurotransmitter noradrenaline, also known as norepinephrine, which causes quiescent melanocyte stem cells to proliferate rapidly, followed by their permanent depletion from the niche Interestingly, Jumpman finds that this is the same pathway that activates hair bulge stem cells, hypothesizing a potential reason for why some grayers are more resistant to balding
Scalp Tissue Analysis: Jumpman’s latest scalp tissue analysis finds that the most significant targets besides the AR are TWIST1/2, PAX1, and AUTS2
Jumpman on Anagen Initiation versus Anagen Prolongation: The signaling required to initiate anagen are different from those required to maintain anagen–PRLR expression inhibition, for example, only maintains anagen. Most people are focused on initiating anagen (hair follicle stem cell activation), whereas Jumpman believes we should focus more on anagen prolongation by targeting transit amplifying cell (TAC) commitment in our hair germs
Reviewed by Ralf: Ralf dropped his Review of PAI-1 inhibitors, C3A, CosmeRNA, and PRLR
Jumpman on Ralf: Jumpman pointed out that Ralf likes thyroid receptors, SFRP1, and mast cells as targets
Jumpman’s Twist-1 Drop: Jumpman dropped a seminal Twist-1 paper, highlighting the target’s history in baldness and the pathways underlying it Please find our notes on the paper here
Inner Root Sheath : The IRS consists of three layers: the cuticle, Huxley’s layer, and Henle’s layer What are the distinct characteristics of each region?
Body Hair: Endana asked if we should try to figure out how to turn body hair into scalp hair, to increase our donor hair supply Herbert thinks the amount of funds required to do so would also enable us to epigenetically reprogram our scalp hair cells to become resistant to DHT.
Bald Countries : Dnovo shared the baldest countries. Can we look at genes more correlated with those from the West and East, to better understand the genetics underlying hair loss?
Smoking: Jumpman informed us that smoking increases prolactin, likely having an effect in our hair loss
5ar Inhibition: VforVegeta warned against long-term 5ar inhibition’s effects on our liver and metabolism It's extremely effective and safe in the short term, but there are not many people who've been taking it for 20+ years, he warned Jumpman and Pevjo identified why 5ar inhibition doesn’t work for many.
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Monoclonal antibodies: Herbert told us how to bring down the cost of monoclonal antibody treatments. Good to keep an eye on, especially if the HMI-115 Phase 1 results come out positive
Hairy Similar Cell Models: Farnsworth hypothesized which cell lines are most similar to hair follicles He likes pancreatic stem cells due to their hypoxic environment, but he wants us to standardize a model across all cells.
Dr. Claire Higgins: We had a great call with Dr Claire Higgins, whose lab at the Imperial College of London is at the forefront of hair and skin research. Dr. Higgins is as imaginative as she is productive–not only did she find that SFRP2 upregulation is a desirable effect for our hair regrowth in the presence of DHT, but she also poses, “Can changing faces save hair loss?”
Research Task: Design a drug that targets FGF5 without inhibiting FGF5s.
Research Task: Design a drug that upregulates RUNX2 without altering RUNX3.
Research Task: What actor would we look like if we downregulated IRF1/4?
Hair Cuts
**Unless stated otherwise, all tokens awarded in this edition of HAIR CUTS will be vested over a two-year period and will also be locked for one year. If you do not claim your HAIR within 3 months of this publication by posting your wallet address in the #hair-drop channel of our Discord server, your right to HAIR will be forfeited
● Jumpman: 5,266 71 Our king reigns supreme as HairDAO’s top researcher Not only did he provide us with his comprehensive analysis of over 50 gene targets, but he also remained a consistent voice of reason on all hairy matters scientifically and operationally for HairDAO.
● Herbert: 4,107.72. Herbert dedicated himself to understanding all there is to know about delivery vehicle research Drawing a comparison to Oppenheimer, our “Father of the Delivery Vehicle Study” spent countless hours on research to orchestrate our next study.
● Endana: 3,027 43 Chicago’s very own Wolf of Wall Street connected HairDAO with over 25 delivery vehicle researchers, introduced us to Dr Bloxham and Dr Powell who are now running our T34 observational study, and continues to provide invaluable operational support
● MTX STUDIO: 2,069 71 Unvested, locked up until 6/30/2024 The incredible team at MTX developed Ready PlayHAIR One. The first of many products with this �� team.
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● JesseH: 2,017 84 Unvested, locked up until 7/1/2024 Jesse acted as HairDAO’s lawyer during our latest fundraising round. He structured our off-chain entities quickly and at a fraction of the cost of most lawyers
● Benjelz: 1,114.95. Benjelz has been a dedicated contributor to HairDAO for over a year now and, more recently, has been instrumental in sourcing exciting compounds which may bind to our top target genes
● CameronS: 360.89. CameronS handled reachout and business development with over 700 chemical labs and compounding pharmacies Ultimately, his work allowed us to find the compounding pharmacy who will produce T34
● Pevjo: 359.10. Pevjo shares valuable tales from his exploratory biohacking ventures, including insights which have helped design our T34 study He also contributed significant research on HDAC inhibitors, Cyclosporine-A, Dutasteride, and numerous other treatments and targets.
● Farnsworth: 324 10 Farnsworth is quite the talented researcher He led the way on researching the roles of EGFR, ERK, miR-221, NOX, HIF-1α, and more.
● Follicle Thought: 278 31 One of the best bloggers in the game, Follicle Thought continues to connect us to hair loss innovators, two of whom are participating in our delivery vehicle study.
● Visangle: 263 30 Visangle is a top biohacker with T levels that would shame a bull shark He contributed research on a wide range of targets, treatments, and topics, and even submitted an article to this edition of Hair Cuts on Androgen Receptor inhibitors. Contact him for a Sneak Peak of Hair Cuts 5
● Eldarlmari: 247.44. A retired gigabrain entered the chat. Eldarmari contributed prominent research on FoxA2, Shh, and epithelial-mesenchymal communication.
● Kabab: 228 23 Kabab transcribed the contact information for over 200 research chemical labs to help us build our supply chain. He also took the lead on building out our Indian network of physicians and compounding pharmacies
● catBellyRubs: 226 23 One of our friendly neighborhood cats, catBellyRubs shared research on DHT and dutasteride, advised us on T34 supply chain, and put HairDAO in touch with Verteporfin-icon Dr Michael Longaker
● Zero: 209 24 Zero, also known as Alphamedian, provided meaningful feedback on our delivery vehicle and T34 studies, analyses of numerous gene targets, and well-thought operational advice
● Fka olive [memo]: 186.22. Memo provided invaluable insight into the tests we are requiring of patients in the T34 trial. He even went as far as developing two specialized diagnostics with Marek Health for the trial
● GorillaHead: 182.80. GorillaHead contributed unique research on a plethora of treatments, including Hydrocortisone, Melatonin, Osteopontin, and Cyclosporine-A
● dNovo24: 174 59 The other of our friendly neighborhood cats, dNovo24 is always there to break the ice with a joke but, more importantly, is putting in the work to become a serious hair loss researcher and shared several valuable studies and insights to help create Hair Cuts 4
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● Copernacus: 152 34 Copernacus opined on a range of treatment options, including HMI-115, Melatonin, Oral Minoxidil, Caffeine, and more. He may have also recruited a patient or two for our T34 observational study
● TheBest: 139.29. TheBest contributed to our research on Twist-1, PAI-1, and today’s most trending treatments, including HMI-115, RU58841, and more.
● Pubehead: 131 43 Pubehead provided significant research on numerous pathways and targets, including HIF-1α, EGLNs, Zyxin, and more in this epoch.
● TCP: 126 43 TCP contributed to our research on Estrogen Receptors, HIF-1α, PKC inhibition, and was always game to find a relevant paper
● potential Dingo: 113.34. potential Dingo analyzed targets, updated us on hair loss companies, and provided operational support He also chipped in with Kabab to grow our Indian network
● Romeo: 101.72. Our in-house data scientist consistently lent his valuable perspectives on a range of topics
● Moemanthe2nd: 96.44. Mo’ money, mo’ problems. We had a real life celebrity join the community to drop some hair loss alpha The community is beyond thrilled to welcome MoeMan to HairDAO
● Kranz: 88.91. Kranz is an absolute marketing whiz, who has taken over HairDAO’s TikTok account, writing several scripts and editing as many videos Last we checked, his invite links were responsible for over 60 new HairDAO members
● Arslan: 81.69. Arslan was able to provide great assistance and feedback for newcomers considering oral dutasteride, oral minoxidil and other treatments
● Guillermo: 70.23. Guillermo provided invaluable direction and connections in setting up our American supply chain.
● Thomasb: 67 31 Thomasb shared valuable information from his gyno experience from oral dutasteride. He also contributed to our research on PRLR, PAI-1, and several additional targets and treatments
● Salvation: 62 76 Salvation provided significant improvements to delivery vehicle study design. He also pondered reproductive health drugs’ benefits to side effects generated by hair loss drugs
● Metabobbly: 61 32 Metabobbly provided continuous updates of his biohacking journey, with a focus on HDAC inhibitors, caffeine, melatonin, lithium, JAK inhibitors, and more.
● Aube Remix: 58 57 Aube Remix was full of tales from his trials and tribulations on dutasteride. He also provided valuable insight on the dutasteride dosage we should use for our delivery vehicle study.
● Blackhole: 57 64 Blackhole contributed to our research on HDAC inhibitors and Minoxidil this epoch.
● Ryaan: 56 69 Our very own inside man at the NIH, Ryaan spent time debating finerenone versus eplerenone He added additional research on 5ar inhibitors and their side effects.
● Vforvegeta: 52 97 Vforvegeta contributed to research on several targets and treatments, focusing on 5ar inhibitors and Ketoconazole Is the itch caused by DHT or something else?
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● Fgsfds: 52 15 Our gigachad referred us to multiple compounding pharmacies and provided research on several targets.
● Aspect x: 47 78 If passion for hair loss had a mascot, it would be Aspect He shared several personal anecdotes and asks the right questions. We have high hopes for Aspect. Now if we could just channel his energy…
● Emmzii: 38 02 Emmzii designed the Hair Inc seal on short notice and posted some high-quality content on social media platforms.
● Officialb: 37 05 Officialb has a wild sense of humor and churned out a meme a minute for several months
● Eleanora caleistah: 36.43. Eleanora made numerous videos and memes, drafted a handful of emails, and provided insights on multiple products for the HairForce
● Recovery Squad: 35 per member Johannes Weniger, Alex Dobrin, and Schmakofant secured our data from our old server in Richard Dunlap’s Discord attack, with us losing only 9 days of data
● 46 members joined the 25 and below club:
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That concludes this Hair Cut, co-created by 87 and counting anons on the internet The HairForce already has a few product drops ready for Hair Cuts 5, including decentralized science’s first human in vivo study, the launch of Ready PlayHair One, and more to be revealed
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