The Group Buy to End All Group Buys: Twist1 Inhibitor

pegasus2

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Edit: Price is $120/g. MOQ 1g.

There are two recently discovered, potent, small-molecule Twist1 inhibitors.

Twist1 deletion keeps murine hair follicles in anagen indefinitely, and a recent study shows that it's dispensible in human DP cells. Twist1 is critical during embryogenesis, but in adults its main functions seem to be causing cancer and preventing hair growth. Twist1 inhibition should be very safe. The safety question with these compounds is potential off-target effects, as with all untested compounds.

On P23, the Twist1f/f hair follicles had just started the anagen phase of the second hair cycle. This anagen phase was sustained to approximately P40, and then was followed by the catagen phase observed at P45 and the telogen phase observed at P54 and P61 (Figure 5A). In contrast, the Twist1Δ/Δ hair follicles always remained in the anagen phase, as histologically examined on P21, P23, P29, P34, P37, P40, P45, P54, and P61 (Figure 5A). A schematic comparison of these differences in the hair follicle growth cycle is presented in Figure 5B. These results clearly indicate that Twist1 plays an essential role in maintaining the hair growth cycle by converting the anagen phase into the catagen phase, which is then followed by the telogen phase. The loss of Twist1 function keeps the hair growth cycle at its anagen phase in adult mice....
These results further support the notion that inducible knockout of Twist1 in young mice leads to arrest of the hair follicle cycle at the anagen phase in adulthood.
Our data also show that the Twist1 protein level in DP cells does not change significantly at different phases of the hair follicle, suggesting that the function but not the expression level of Twist1 is regulated during the hair growth cycle.
The function of Twist1 is altered by dimer composition having preference for different E-box motifs, and differentially regulating FGF signaling.

Inducible Knockout of Twist1 in Young and Adult Mice Prolongs Hair Growth Cycle and Has Mild Effects on General Health, Supporting Twist1 as a Preferential Cancer Target

R-spondin is what keeps hair in anagen, and its levels are regulated by FGF signaling. Silencing FGFR1/2 keeps R-spondin secretion high and prolongs anagen significantly, resulting in these long-haired mice:
fgfrnull.JPG

Fgf and Wnt signaling interaction in the mesenchymal niche regulates the murine hair cycle clock

We propose that the DP secretes R-spondins to synchronously instruct activation of bulge/hair germ progenitors and neighboring hfDSCs in order to enable coordinated HF regeneration.
Transcriptional Profiling of the Adult Hair Follicle Mesenchyme Reveals R-spondin as a Novel Regulator of Dermal Progenitor Function

R-spondins combine with LGR4/5/6 to prevent degradation of Wnt receptors. This is required to keep Wnt signaling and the anagen phase going.
The R-spondin/Lgr5/Rnf43 module: regulator of Wnt signal strength
Stem Cell Intrinsic Mechanisms Regulating Adult Hair Follicle Homeostasis

Altogether these findings suggest that a shift in the balance between Twist homodimers and heterodimers is what induces FGFR to inhibit r-spondin expression. The balance between Twist1 dimers could be altered by BMPs, TGFbeta, an increase in Twist1 expression, or a reduction in TCF proteins.

Disease-causing mutations in TWIST1 can impact dimer formation or shift the balance of different types of TWIST1 dimers in the cell, which may underpin the defective differentiation of the craniofacial mesenchyme. Functional analyses of the loss and gain of TWIST1–E-protein dimer activity have revealed previously unappreciated roles in guiding lineage differentiation of embryonic stem cells: TWIST1–E-protein heterodimers activate the differentiation of mesoderm and neural crest cells, which is accompanied by the epithelial-to-mesenchymal transition. At the same time, TWIST1 homodimers maintain the stem cells in a progenitor state and block entry to the endoderm lineage.
Previous studies have highlighted the differential functions of TWIST1 dimers in the osteogenic differentiation of the cranial sutural mesenchyme (21, 26), which is mediated by their targeted action on fibroblast growth factor (FGF) signaling (25, 27, 28). For example, the TWIST1-TCF3 heterodimer promotes mesenchymal stem cell (MSC) proliferation, while the TWIST1 homodimer activates FGFR2, OCN, and BSP expression for ossification.
the homodimer and the heterodimer antagonistically regulated a subset of critical developmental regulators such as those associated with TGF-β signaling (e.g., Tgfbr1 and Pdgfrα) and WNT signaling (Prickle1 and Dkk1) (Fig. 6C). Other genes, represented by Fgfr2, Jun, Chd7, and Mmp2, were activated more effectively by T12 than by TT
a.JPG

1657506901703.png

TWIST1 Homodimers and Heterodimers Orchestrate Lineage-Specific Differentiation


TT is Twist1 homodimers, and T12 is Twist1/TCF12 heterodimers. As the balance shifts to significantly more homodimers Wnt signaling is repressed. It is also repressed slightly when heterodimers are highly overexpressed, and EMT increases sharply which will also cause hair loss. All Twist1 expression upregulated FGFR, but Twist heterodimers upregulated it the most. This contradicts the other study, but it seems likely that shifting the balance towards Twist1 homodimers would be good for us. The balance would have to be highly skewed towards T12 heterodimers in prostate cancer to sharply upregulate EMT. It seems likely that it's also skewed in that direction in Androgenetic Alopecia. SCUBE3 or Cyclosproine A should shift that balance back towards homodimers, but eliminating Twist1 altogether would be even better.

Twist1 is upregulated in balding scalp DPCs. It's also upregulated in another androgen-dependent disease, prostate cancer, via NF-κB and ETV1. It induces aberrant activation of AR, driving prostate cancer migration and resistance to androgen deprivation therapy.
we found TWIST1, a DP signature gene expressed in both BAB and BAN (Figure 1e and see Supplementary Table S1a), being up-regulated in BAB[balding men] (Table 1). Twist1, a basic helix-loop-helix protein, is crucial for anagen-to-catagen transition during the hair growth cycle with Twist1 protein ablation in adult mice DP resulting in prolonged anagen (Xu et al., 2013). Hence, TWIST1 up-regulation in DPCs of balding scalps compared to non-balding scalps may result in accelerated transition from anagen to catagen and thus a shortened period of anagen during the hair cycle, a phenomenon in balding scalps that leads to the formation of short vellus hairs instead of long terminal hairs (Paus and Cotsarelis, 1999).
TWIST1 also interacts and binds to HDAC4 (Danciu and Whitman, 2009) to regulate gene expression (Gong and Li, 2002, Lee et al., 2003). In addition, binding of TWIST1 at E-boxes in the AR promoter region results in up-regulated AR expression

Differential Expression between Human Dermal Papilla Cells from Balding and Non-Balding Scalps Reveals New Candidate Genes for Androgenetic Alopecia

inhibition of AR signaling by androgen depletion and the novel antiandrogen enzalutamide induced PKC and RelA activation, resulting in Twist1/AR induction at the transcript level. Moreover, inhibition of NF-κB signaling prevented enzalutamide-induced Twist1 and AR induction. Finally, NF-κB was activated in both castration-resistant and enzalutamide-resistant cells. In conclusion, NF-κB signaling was responsible for Twist1 upregulation by PKC in response to AR inhibition, resulting in aberrant activation of AR. NF-κB signaling thus appears to play a critical role in promoting both castration resistance and enzalutamide resistance in PKC/Twist1 signaling in prostate cancer.
our results clearly showed that androgen deprivation increased the ROS levels in PCa cells, and this effect was partially abolished by the free-radical scavenger NAC. These findings seem to be inconsistent with previous reports that androgen signaling increases oxidative stress (Ripple et al., 1997; Pinthus et al., 2007; Pathak et al., 2008). However, these studies were conducted using overdoses of androgen beyond physiological levels. Our results are supported by previous observations of increased oxidative damage to cellular molecules with the development of malig- nancies (Bostwick et al., 2000; Oberley et al., 2000) and aging (Ghatak and Ho, 1996; Lu and Finkel, 2008; Maynard et al., 2009), accompanied by declining testosterone levels. Furthermore, our results are sup- ported by the finding that ADT for PCa may increase the risk of death from cardiovascular disease, which is closely implicated in oxidative stress (Hakimian et al., 2008). It has also been shown that increased oxidative stress in rats after castration results from dramatic increases in ROS-generating NAD(P)H oxidases and significant reductions in ROS-detoxifying enzymes (Tamm et al., 2003). In addition, the major ROS scavenger MnSOD (mitochondrial superoxide dismu- tase-2) shows decreased mRNA levels in PCa after ADT (Best et al., 2005), and oxidative stress-related genes, including thioredoxin, peroxiredoxin 5 and MnSOD, are reduced in the rat prostate after castration (Pang et al., 2002). Taken together, these findings may indicate that both androgen deprivation and overload can increase oxidative stress
Castration resistance of prostate cancer cells caused by castration-induced oxidative stress through Twist1 and androgen receptor overexpression
Twist1 depletion strongly repressed the migration of prostate cancer cells, comparable in effect to AR knockdown
In this study we demonstrate that androgen-activated AR promotes the expression of Twist1 in prostate cancer cells. Since there is no evidence for AR binding to the Twist1 promoter, we hypothesized that the AR effect on Twist1 expression is indirect and mediated by a direct target gene of AR. We provide evidence that the Ets protein ETV1, whose gene is a direct target of AR, mediates the AR positive effect on Twist1.
Khatiwada, Prabesh et al. “Androgen up-regulation of Twist1 gene expression is mediated by ETV1.”

These are small-molecule Twist1 inhibitors:

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A couple other interesting graphs here from the Twist1 dimer study. This shows Twist1 deletion(red bar) increases Wnt receptor Fzd10. This suggests that Twist1 deletion is increasing R-spondin secretion. One of the most highly upregulated genes by Twist1 is Sox9, which stops Wnt signaling and is upregulated just before catagen. Not incidentally, it regulates and is regulated by FGFR.

our results demonstrate that organ fate commitment and progenitor cell expansion are coordinately controlled by the activity of a Sox9/Fgf10/Fgfr2b feed-forward loop in the pancreatic niche. This self-promoting Sox9/Fgf10/Fgfr2b loop may regulate cell identity and organ size in a broad spectrum of developmental and regenerative contexts

@EndlessPossibilities found that a rare disorder called Saethre Chotzen syndrome, which is caused by a mutation or deletion in Twist1 resulting in severely impaired Twist1 signaling, also results in a lower hairline and these people don't seem to suffer from alopecia. Although this is very anecdotal with a small sample size, it suggests that Twist1 inhibition might be enough to prevent Androgenetic Alopecia. Even if this is true that doesn't mean it's enough to reverse it, but it would be one of the keys.

331254_1_En_16_Fig4_HTML.jpgindex.jpg775EBDA5-8C1E-487C-A24B-B2F765C3FE8E.pngAD276F71-66B0-4008-A49E-99DDA0FB7040.png
Given that we have previously established the requirement of TWIST1 for tumorigenesis in oncogene-driven lung cancer with these genetic backgrounds (10), using harmine derivatives may be a viable therapeutic option to treat oncogene-driven NSCLC both in the treatment-naïve and acquired resistance setting. Additionally, as TWIST1 is rarely expressed post-natally (45,46), pharmacological inhibition of TWIST1 may be associated with minimal side-effects.

The two compounds for Twist1 inhibition are harmine derivative 19a, and ADQ. Harmine comes from the plant Syrian Rue, which is a folk remedy for hair loss. Harmine preferentially inhibts Twist1/e-protein heterodimers. That may make it preferable, but it also inhibits Wnt signaling by inhibiting DYRK1A. It is also a psychedelic and displays neurotoxicity. Due to the more favorable off-target effects the group buy will be for ADQ.
 
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RagnarLothbrok

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Uhm. So you're saying just in a few years CRISPR knocking out Twist1 would pretty much be a cure with little side effects? Sounds too good to be true as usual

Also what happens if your hair is forever in anagen phase, wouldn't you become a werewolf everywhere lmao

Whats the name of the molecules that inhibit Twist1 and what % of it does it inhibit?
 

pegasus2

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Uhm. So you're saying just in a few years CRISPR knocking out Twist1 would pretty much be a cure with little side effects? Sounds too good to be true as usual

Also what happens if your hair is forever in anagen phase, wouldn't you become a werewolf everywhere lmao

Whats the name of the molecules that inhibit Twist1 and what % of it does it inhibit?
You definitely wouldn't want to knock it out with CRISPR everywhere, only in the HF. That would have to be studied a lot further before doing that. It's much better to just inhibit it with these compounds
 

Zon Ama

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I love you guys. Even if I dont understand the science behind all this, I am sure somebody will find the cure with the group buys here

Where is @FollicleGuardian btw? Did he get the cure and left us all alone to suffer?
 

Hairful

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I love you guys. Even if I dont understand the science behind all this, I am sure somebody will find the cure with the group buys here

Where is @FollicleGuardian btw? Did he get the cure and left us all alone to suffer?

Of course you love them. They’re being Guinea pigs for us
 

pegasus2

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But how this will address the DHT issue?
Anti-androgens may always be the first-line defense against hair loss. However, in prostate cancer Twist1 knockout alone was as effective as complete androgen receptor knockdown.
 
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Roeysdomi

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Anti-androgens will always be the first-line defense against hair loss. However, in prostate cancer Twist1 knockout alone was as effective as complete androgen receptor knockdown.
Iirc last time you talked about twist1 in discord there was somthing preventing from us actually using it , i think it was the WNT pathway if im not wrong
 

Redgate

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Isn't ASC-J9 also inhibiting Twist1? Any reason to get on these if already on it (aside from stains)? Can they be combined?
 

pegasus2

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Iirc last time you talked about twist1 in discord there was somthing preventing from us actually using it , i think it was the WNT pathway if im not wrong
The lack of any good compounds to inhibit it.
 

pegasus2

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Isn't ASC-J9 also inhibiting Twist1? Any reason to get on these if already on it (aside from stains)? Can they be combined?
Likely, but not as much as these molecules
 

RagnarLothbrok

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do we have any human data on side-effects rather than that smol mice study? it sounds less scary than Shh pathway so that attracts me, but isn't most potent WNT upregulators cancer related?
 

pegasus2

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do we have any human data on side-effects rather than that smol mice study? it sounds less scary than Shh pathway so that attracts me, but isn't most potent WNT upregulators cancer related?
There is no chance that inhibiting Twist can cause cancer. Twist1 promotes tumor growth, and these two drugs were developed to treat cancer. It will only upregulate R-spondin in tissues where it is supposed to be upregulated, like the hair follicle. Even in prostate cancer RSPO is downregulated, and lower R-spondin levels are associated with lower survival rates. Twist1 activates Wnt target genes also, differentially from beta-catenin. It shifts the pathway towards EMT which promotes tumor progression.
 
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nimdok

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Great finds, don’t wanna get my hopes up too much but this sounds promising
 
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