Hair Cuts 2

Hair Cuts #2 (February 25, 2023 - April 7, 2023) ____________________________________________________________________________ **Potential future areas of study have been bolded.

HairForce! Welcome to our second issue of HairDAO Monthly’s, where you can get your hairy alpha. These documents stitch together the most important conversations and happenings within the DAO, providing a compressed recap of what you may have missed while you were off losing your hair. As our treasury now sits at $450k, we’ll have the opportunity to translate our best ideas into real research and intellectual property (IP) over the next several months.

Highlight: Delivery Vehicles Synthetic Pug summarized the importance of investigating delivery vehicles further - not only could better solutions enhance the bioavailability of existing treatments to our follicles, but their increased specificity could also limit the side effects of ALL proposed hair loss treatments, increasing the therapeutic window. However, as we learned from Ralf, it’s no simple task to administer an active pharmaceutical ingredient (API) to the correct cells in a human hair follicle.

While we apply topical solutions to the skin, we need the API to travel alongside the hair follicle’s surface through the entire epidermis, reaching the basement membrane–where the epidermis and dermis meet and the hair bulb resides. Ralf believes that in order to truly stimulate hair growth, we need to focus on the mesenchymal command center, which includes both the dermal papilla and dermal sheath cells located at the base of the hair bulb. Kevin McElwee chimed in with a pseudo-agreement to Ralf’s claim, posing that the cup cells in the dermal sheath should be our primary focus over the dermal papilla. No matter the hair bulb mesenchyme group we target, a problem arises: blood vessels are attached to the entire hair bulb. Once an API arrives to the region, it’s highly likely to go systemic through our blood supply. We’re thus left with the following challenge: to get drugs through the entire epidermal layer of our skin such that it accumulates in the hair bulb, without absorption by surrounding blood vessels and skin tissue.

A newcomer to our community, Herbert led our efforts to find better solutions. He drafted his own 20 page literature review of today’s most innovative vehicles, while also submitting a corresponding study to the HAIRy Research Bounty. Herbert’s study plans to source multiple

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nanoparticle vehicles from respected researchers, followed by testing each vehicle’s efficacy and tolerability on human scalp skin in a Franz diffusion cell model. The DAO is currently executing Herbert’s first step – pre-selected researcher reach-out – and is already in touch with several prominent delivery researchers. Despite a lack of assistance from traditional scientists thus far, Herbert continued to make strides in his own vehicle selection, finding that polyactic acid (PLA), polycaprolacton (PLC) and polystrene nanoparticles have the highest likelihood of successfully reaching the bulb. Dukenuke chimed in with ionic liquids as a competing candidate. Jumpman posited how to reach the dermal papilla in bald skin, where only a vellus, miniaturized follicle remains. While the discussion remains unfinished, we’ve made great strides this month, which will surely result in a new study design.

Conversations: This month’s additional conversations outlined much more than what is listed below, only highlighted due to their potential wide ranging effects.

PAI-1 Stays Hot Jumpman and Pubehead continued to unveil new, bullish information about PAI-1. Some of the newly found benefits of PAI-1 inhibition include increases to the proangiogenic factors FGF-2, VEGF-A, KitL, and G-CSF. Synthetic Pug pondered - because DHT acting on androgen receptors (AR) in the dermal papilla triggers the secretion of too much TGFb, which then triggers PAI-1, does that mean AR activity is indirectly involved in activating PAI-1? Well, it appears that researchers are currently stumped. Jumpman theorized that PAI-1 activation downstream of AR may be dependent on SMAD or Rho/Rock pathways. Uncovering the complete AR - PAI-1 pathway may one day make for an excellent study design.

PAMP OUR IP - Jumpman boosted the valuation of our T3/T4 study significantly, finding that T3/T4 may inhibit Pi3k, PAI-1, and Twist-1 signaling, three very desirable traits of hair loss treatments. Pi3K/Akt pathway inhibition fully reversed AGA in SCD1 knock-out mice, another target recently discovered by the Jump. Inhibition of the Pi3k/Akt pathway is proposed to reverse AGA by turning off TGF-beta-induced PAI-1 and resultant Twist-1 expression. It now looks like HairDAO’s Thyroid Hormone treatment could inhibit all three AGA-promoting pathways.

Angiogenesis and Minoxidil’s MoA Does increased blood flow improve hair loss? A popular theory, Dxdy1104 suggested that we might all want to start taking cold showers, as the shock from the cold water puts the circulatory system into overdrive, driving more nutrients to our hair bulbs. Unfortunately, the community was not sold on the idea, sharing pictures of Wim Hof and the bald uakari. More interesting than Wim Hof, the bald uakari monkey is completely bald despite so much blood flow to its head that its face is red. Community members persuasively argued that androgenetic alopecia is more likely driven by stem cell defects and loss of cellular adhesion and signaling, rather than decreased blood flow.

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Despite having been able to read our public discourse, Andrew Huberman stated on his most recent podcast that increased blood flow is Minoxidil’s primary mechanism of action. However, alongside angiogenesis, Jumpman pointed out that Minoxidil inhibits GSK3b and SFRP1, upregulates LHX2, and promotes keratinocyte differentiation - mostly mediated through the upregulation of Hepatocyte Growth Factor (HGF), an executor pathway outlined by Ralf as good for hair growth. Another area for potential study arose from this conversation - Keratinocyte Growth Factor (KGF/FGF7) is upregulated 7-fold in AGA and might induce cellular hyperproliferation, while inhibiting the required cellular differentiation that HGF promotes. Could the ratio of KGF/HGF have a role to play in AGA? We’ll need to generate some new IP to know. While angiogenesis may help, Minoxidil’s exact mechanism of action remains unknown, serving as yet another example of how little we truly know about hair.

FGF, or Why Too Much Proliferation is Bad MolotovRibbentrot furthered the discussion on fibroblast growth factor (FGF), another executor pathway referenced by Ralf. Synthetic Pug outlined the dichotomy of different FGF isoforms in hair loss: the pro-anagen isoforms appear to be upregulated by Wnt and Shh signaling, while the pro-catagen isoforms appear to be upregulated at the end of anagen. Jumpman jumped into the conversation by pointing out that FGF5 (and potentially KGF/FGF7) inhibits cell differentiation by promoting hyperproliferation. Too much cellular proliferation is bad, if hair follicle epithelial stem cells do not have time to divide.

Jumpman then provided an example from Angela Christiano, where she showed that Sox9 repression by Trps1 led to hyperproliferation, which caused hair loss. In a separate conversation about the role of hyperproliferation in hair loss, Herbert also shared that continuous Wnt/B-catenin activation (hyperproliferation) can trigger senescence, also associated with hair loss. The conversation culminated in the community suggesting we further study Trps1, Lhx2, Tnc, and SERPINE1.

Early-stage Targets: Our conversations this month yielded several targets likely play a meaningful role in hair loss: ●

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Cyr61, Wif1, and Frzb - Researchers at the University of Calgary found that Cry61, Wif1, and Frzb were all overexpressed at the beginning of the hair follicle’s development, when dermal papilla cell specification begins. Dermal papilla cell differentiation from its parent dermal sheath fibroblasts is vital for hair regrowth, as previously outlined by Kevin McElwee.

SCD1 - SCD1 is critical in maintaining the hair epithelial stem cell niche through the stabilization of hemidesmosomes. Jumpman found that PRL inhibition, Hope Medicine’s mechanism of action, likely leads to SCD1 upregulation. Can we upregulate SCD1, to achieve a similar effect as Hope Medicine’s PRLR-antibody treatment?

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SMAD3 - We discussed SMAD proteins earlier, as they may determine PAI-1 expression downstream of androgen receptor activity. SMAD3 returned later on, as researchers were able to restore hair bulb size in mice by delivering miR-122-5p in exosomes to the mice’s follicles, which downregulated SMAD3.

TRH-Associated Genes - Thyrotropin-releasing hormone (TRH) promotes mitochondrial functioning and perinuclear biogenesis, simultaneously increasing transcription of reactive-oxygen species (ROS) scavengers - such as catalase and superoxide dismutase 2 - and thus reducing ROS accordingly. It was found that KRTHA1, KRTHB6, KRT6B, MGC10814, UBCE71P5, and KRTHA5 were downregulated over two-fold, while the gene NEF3 was upregulated over two-fold after TRH exposure.

NRF2 - Synthetic Pug found that the gene upregulates antioxidant activity, VEGF, and IGF-1. NRF2 is upregulated by C3A, furthering our interest in the natural compound.

FOX01 - Jumpman found that FOX01 suppresses androgen receptor activity, but only in the presence of DHT. Could FOX01 upregulation substitute 5ar-inhibition in our hair stacks?

Speaking of Jumpman, our most prolific researcher isn’t keen on IGF-1 as a treatment, due to it being upregulated in balding vertex scalps. On the other hand, IFGBP3 is downregulated about 18 fold in AGA. The divergence of opinions on IGF-1 highlights the need for further research into the target.

GPC4 and GPC6 - Rare in androgenetic alopecia research, both glypicans’ expression in different hair cell zones were analyzed at all three stages of the hair follicle cycle.

Interestingly, GPC6 upregulation during non-anagen could be due to it causing hair growth at those times, but for the next follicle. Should we alter our hair cycle timescale, trying to upregulate factors found in telogen, as opposed to anagen?

SASP genes - Synthetic Pug shared a list of SASP genes that we may want to downregulate, as they likely cause cellular senescence in cells both inside and surrounding the hair follicle.

PER2 - The long-forgotten gene may be worth looking back into, as it regulates PAI-1 promotion. Jumpman and Pubehead form quite the tandem when going into the big brain details upstream and downstream of PAI-1.

Pending Studies: The HAIRy Research bounty has commenced. We are now fielding both studies and equity investments for our second funding round. In turn, both HairDAO’s internet-native and traditional scientists have begun submitting study proposals: ●

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Verteporfin - by Endana. Endana generated the first submission for our next investment, proposing a pay-for-success reservation system to accelerate Dr. Bargouthi’s testing of verteporfin in hair transplants. Due to feedback from the community, Endana’s proposal now includes a potential equity investment into Dr. B’s practice.

Delivery Vehicles - by Herbert. As discussed, the creation of a vehicle that can be delivered to the base of the hair follicle without systemic absorption would dramatically improve the viability of all existing hair loss drugs, even those with significant side

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effects. We’d first identify as many promising delivery vehicles as possible, and then test them on human scalp skin mounted to a Franz diffusion cell model. The top-performing vehicles would advance to in vivo trials, either on human scalp skin on murine back or directly in humans, depending on regulatory hurdles.

TRH (awaiting finalization) - by Ralf. Thyrotropin-releasing hormone (TRH) has prolonged anagen, upregulated melanin synthesis (hair color), and improved mitochondrial functioning and biogenesis in the human hair follicle. The present study builds directly on Paus et. al’s 2010 study, in which human hair follicles were treated with TRH in organ culture. In the prior study, numerous hair growth-promoting results were shown in the treated follicle group: anagen was significantly prolonged; the catagen-promoting effects of TGFb2 were substantially negated; and Ki-67+ matrix keratinocytes were prominently upregulated.

TRMP5 & TAS2R4 (awaiting finalization) - by Ralf. In a human hair follicle organ culture, transient receptor potential cation channel subfamily M member 5 (TRPM5) stimulation was shown to prolong anagen, upregulate key hair anagen-promoting growth factors (such as those in the overarching WNT pathway), and downregulate catagen-inducing TGF-b2. Additionally, Paus et. al recently discovered that TAS2R4 activation inhibits human scalp hair follicle growth in organ culture. The combination of TRPM5 agonist and a TAS2R4 antagonist may make for an effective chemosensory approach to slowing hair loss.

FK-506 (awaiting finalization) - by Ralf. Two immunosuppressive drugs, FK506 and Cyclosporine-A (CsA), have both exhibited hypertrichotic effects in their non-androgenetic alopecia applications. While CsA has shown promising hair growth data, the drug is too toxic to be used as a treatment. However, FK-506’s lower toxicity profile and rare topical-only effects render it an excellent candidate to test using our human scalp skin organ culture model. Ralf wants to test for key transcriptomic differences enacted by FK506, to generate new lead compounds to act on the same pathways.

Stemson Therapeutics (awaiting finalization) - by Geoff Hamilton. Stemson Therapeutics utilizes induced pluripotent stem cells (iPSCs) to generate and transplant human scalp hair follicles de novo. The company most recently demonstrated that their de novo human hair follicles could be successfully transplanted and grown directly in immuno-compromised mice. Stemson is now raising $20mn USD to prove that their follicles can be successfully implanted and grown in human skin tissue on the backs of mice, their final step before human clinical trials.

HairClone (awaiting finalization) - by Jennifer Dillon. HairClone seeks to treat hair loss by extracting, culturing, and then re-administering dermal papilla cells from the occipital region of the scalp, where our hairs remain unaffected by androgens. Currently, HairClone is only licensed to administer its cell therapy treatment in the United Kingdom (UK). The company is seeking a $500,000 investment to collect proof of concept in-human data, such that they can fine-tune their protocol for Phase 1 clinical trials and multi-country expansion.

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Treatment Reviews: ●

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Melatonin lipid nanoparticles generated over an 85% increase in hair width over three months, per our resident Macaque. Herbert and Synthetic Pug then debated its mechanism of action, which remains an open question.

Synthetic Pug found that Ginkgo Biloba may be beneficial through mesotherapy, as it has an inhibitory effect on FGF5, which prevents the hair cycle from restarting.

Why do 5-alpha-reductase (5-AR) inhibitors stop working after 5-10 years? Jumpman states the best answer we’ve heard to date.

After questioning by Gurgeh, Fgsfds provided an excellent review of Viviscal & Nourkrin as an effective solution for hair loss. Check out tilapia skin grafts, WILD! CaptainSua & Hoor warn there may be long-term side effects.

Herbert and DukeNuke informed us that, if you don’t respond to off-the-shelf Minoxidil, Minoxidil Sulfate may prove more effective. Benjels is either a non-responder to oral minoxidil, or he’s experiencing some bad shedding.

BenjamTD and Fka Olive had a fantastic discussion on how HCG and BPC-157 might . help cure post-finasteride syndrome. BenjamTD added a little LSD to his protocol Fka Olive has seen >2000 patients on thyroid hormone replacement therapy. He believes that the treatment is more likely to work on diffuse thinners than patients experiencing hairline recession.

Synthetic Pug shared a study suggesting that Arctiin might be an effective antioxidant to combat cellular senescence in the hair follicle.

Metabobbly has seen some minor improvements from red light therapy, when using a very strong light. Averbs123 thinks red light is useless, although efficacy may be determined by the strength of the wavelengths.

Charging01 asked - Has anybody tried PTD-DBM combined with microneedling?

BenjamTD did, but with no results.

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PAI-1, C3A, CosmeRNA’s androgen receptor inhibitor, and Hope Medicine’s PRLR antibody remained the community’s most discussed pharmaceutical topics, culminating in our first Reviewed by Ralf segment. The document was heavily edited by Jumpman, Synthetic Pug, Endana, and Eleanora. Ralf was busy with University of Miami obligations and then fell sick, but will hopefully be back on the horse soon. A world with a healthy Paus is a far better world.

Hairy Matters 2 came out. In it, Ralf discussed basic hair biology, epithelial-mesenchymal pathway communication, the hair follicle’s chemosensory receptors, neuroendocrinology’s relation to hair, and the immune system’s role in hair loss.

We officially launched the patient portal! Community members can now upload genomic and treatment protocol data in order for us to run decentralized clinical trials, discover new targets, and recommend treatments. Submit your data, earn HAIR!

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On hair loss’s relation to hair graying, the answer remains unknown. Ralf has brought up numerous times that hair rapidly grays before it falls out. On the other hand, prominent melanin researchers, such as Desmond Tobin, believe that gray hair can survive healthily for a long time period. The answer likely depends on the pathways at play, on a per-patient basis.

Jumpman prescribed ‘No Fap’ to the community. Fapping likely accelerates hair loss Does creatine cause hair loss? Synthetic Pug cited research saying the link isn’t that strong, but Jumpman questioned that theory - creatine raises testosterone after all!

CaptainSua wanted insight into using glycerin instead of propylene glycol as a vehicle for his topical minoxidil/finasteride. Unfortunately, glycerin is a poor permeator.

Herbert gave a fantastic overview of Dr. Claire Higgins’s theory of hair follicle miniaturization.

Herbert also came to the conclusion that cyanidins are worth investigating further, due to their ability to translocate membrane androgen receptors. Wisdom of the crowd continues to move nicely for both PAI-1 inhibitors and C3A.

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HAIRy Rewards: **Once received, all HAIRy rewards will be vested monthly over a one year period. ● ●

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Ralf: 730.16, per our contract.

Jumpman: 670. Jumpman continues to rival Ralf as the world’s most prolific hair loss researcher. This month, he both ideated and analyzed a plethora of targets, including PAI-1, TGFb, Twist-1, FGFs, Scd1, Fox01, GPC4 and 6, and PER2. The entire hair loss community is grateful for Jumpman’s pursuit of excellence.

Herbert: 380. While a newcomer to our community, Herbert led our delivery vehicle research. His work culminated in an ongoing study design, which galvanized HairDAO to get in contact with several leading delivery vehicle researchers. He also contributed novel ideas to the theory of balding - enforcing our belief in the importance of the dermal sheath - and analyzed several treatments and targets along the way.

Synthetic Pug: 320. Synthetic Pug continues to lead our cellular senescence research, epitomized in the C3A section of our upcoming Reviewed by Ralf segment. He also contributed significantly to our debates on NRF2, FGFs, PAI-1, TGF-b, melatonin, and delivery vehicles. It would be no surprise for Synthetic Pug’s research to culminate in a formal study soon enough.

Benjels: 250. Benjels continues to handle both mission-critical operations and research for HairDAO. This month, he played a leading role in the Patient Portal’s launch and the HairDAO Multisig operations, as well as delivery vehicle, PAI-1, and verteporfin research.

Sminepepe: 200. Sminepepe led numerous mission-critical operations for the DAO, putting together presentation materials for our upcoming demo day, editing and creating community-generated content for our social media channels, and moderating our public debate.

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Schmakofant: 180. Schmakofant played a leading role in the launch of the Patient Portal, both writing code and advising Andy1 and Jay1126 on its development. He also launched the HAIR token dashboard on Dune Analytics, set up our voting forum on Snapshot, and deployed HAIR to the Optimism Network.

Endana: 120. Endana completed the first submission to the HAIRy Research Bounty, sourcing and ideating a strategy for HairDAO to support Dr. Barghouthi’s research. He also generated several questions for Ralf on CosmeRNA. DukeNuke: 110. DukeNuke contributed significantly to the debate on HairDAO’s delivery vehicles and meta-design, including feedback and ideas for the Research Portal and testing model trade-offs.

Fka Olive: 100. Fka Olive provided valuable insight on thyroid hormone therapies, additional hormone-dependent treatments, and novel topical formulations.

BenjamTD: 80. A tried and true expert in all current hair loss treatments, BenjamTD contributed significantly to our research on the downstream effects of 5ar inhibition. His performance in our first commercial was also a massive hit.

Fgsfds: 80. Fgfds led our research and analysis on Viviscal, a popular natural treatment.

He also contributed to our discussions on melatonin and hormone therapies.

Johannes Weginer: 50. Johannes provided valuable advice on the Patient Portal and continues to push forward numerous HairTables, such as our Contributor and DeaFlow forms.

Kenichi & Ismail.Malik: 50 each. Our tireless video editors put together several hilarious, informative clips from our longer form content. Both contributors play a critical role in bringing high-level science to the mainstream.

The 20 Club - the following members provided insightful ideas and remarks on targets, treatments, HairDAO’s design, and more: MolotovRibbentrot, Dxdy1104, PubeHead, Metabobbly, Raicas, Romeo, Ryaan, Eleanora, Gurgeh, Hoor, Vincent, Ghter, Pevjo, Kevin McElwee, Ndimension, Medo, Yigitamkk, and CaptainSua.

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That’s all for now, folks. The next Monthly’s is already underway, with both new targets ideated and old targets analyzed. Long Live HairDAO

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