Restoring Wnt/β-catenin Signaling Is A Promising Therapeutic Strategy

[pegasus2]

This is an interesting paper on Wnt signaling. It isn't about hair, but it is relevant to us all the same. Hair loss research might not be hip, but the involved pathways intersect with diseases that are.

Restoring Wnt/β-catenin signaling is a promising therapeutic strategy for Alzheimer’s disease

long-term moderate exercise and environmental enrichment can stimulate Wnt/β-catenin signaling by reducing DKK1 protein levels and increasing LRP6 and Wnt3a protein levels in hippocampus of adult animals [29, 135]. These findings suggest that activation of Wnt/β-catenin signaling is a potential mechanism underlying the cognitive improvement associated with an active lifestyle.

Some people don't want to hear this, but a sedentary lifestyle can only aggravate hair loss. That doesn't mean exercise is going to cure hair loss or prevent it. Far from it. It's a good idea to get off the couch though.

The paper mentions the following DKK1 inhibitors:

estrogen-induced neuroprotection and attenuation of tau phosphorylation are associated with DKK1 inhibition and subsequent activation of Wnt/β-catenin signaling [139]. Together, these findings suggest that inhibition of DKK1 is a potential mechanism for estrogen-induced neuroprotection.

A virtual screen of the National Cancer Institute database for chemical compounds identified a small molecule, IIIC3 (NCI8642, gallocyanine), as a DKK1 inhibitor [143]. IIIC3 can inhibit DKK1 binding to LRP6 with an IC50 of 3 μM [143], and revert DKK1-mediated inhibition of Wnt/β-catenin signalling [143, 144]. Moreover, IIIC3 can reduce basal blood-glucose concentrations and improve glucose tolerance in mice

Curcumin, a natural compound found in the plant turmeric (Curcuma longa), displays protective effects in various animal models of AD [150, 151]. Studies have shown that curcumin can potentially promote Wnt/β-catenin signaling by increasing the expression of Wnt proteins and Wnt co-receptor LRP5/6 and suppressing the expression of Wnt antagonist DKK1

Safety concerns around Wnt agonists:

There is always a concern that overstimulation of Wnt/β-catenin signaling can promote cancer because aberrant activation of Wnt/β-catenin signaling can lead to tumor formation [6, 109]. However, there are no reports of increased incidence of cancer in families carrying LRP5 gain-of-function mutations, and Sost- or Dkk1-deficient animals do not have an increased risk of tumor developments [183]. Nevertheless, the therapeutic application of Wnt activators should be given precisely to restore, but not overactivate, the Wnt/β-catenin signaling pathway in AD patients.

 

[pegasus2]

If anyone is interested in trying gallocyanine (IIIc3) as a dkk1 inhibitor, it's potent, dirt cheap, and soluble in anything. The only problem is that it's not stable in solution so you'll have to make it fresh every day. On the bright side the powder is stable at room temperature for months, so you don't have to take it in and out of the freezer every day. It only takes a couple minutes a day to put a couple ml of minoxidil or stemoxydine in a separate bottle and then shake it up. @whatevr This might be easier for you than making MAP liposomes. It isn't quite as effective a Wnt agonist as WAY-262611, but it's close, and the cost is only around $5/mo. compared with $500/mo. for WAY-262611.

https://dev.biologists.org/content/develop/144/20/3819.full.pdf?with-ds=yes

 

[pegasus2]

Genetic interaction of PGE2 and Wnt signaling regulates developmental specification of stem cells and regeneration

PGE2 regulates wnt activity by direct phosphorylation of β-catenin and GSK3β

Our work shows that PGE2 acts via cAMP/PKA signaling to directly modify β-catenin stability and emphasizes the functional importance and evolutionary conservation of these interactions in organ development and regeneration. PGE2-mediated regulation of Wnt signaling was previously demonstrated in cell lines. The proposed biochemical mechanisms mediating the interaction are remarkably diverse and may depend on the particular cell line studied (Clevers, 2006). Castellone et al. used colon cancer cell lines to demonstrate PGE2/PI3K-mediated activation of Akt, leading to dissociation of Axin1 from the destruction complex (Castellone et al., 2005). While we cannot exclude the existence of this biochemical interaction in vivo, our studies suggest that activation of PKA is the functionally significant effector downstream of PGE2 in HSCs. Likewise, both phosphorylation of GSK3β and β-catenin by PGE2-based activation of PKA have been implicated in Wnt signaling regulation in vitro. Fujino et al. used the transformed HEK293 cell line and showed that both PKA and PI3K could function downstream of PGE2 to phosphorylate GSK3β (Fujino et al., 2002). However, PGE1-induced activation of PKA and phosphorylation of β-catenin at S675 was cell-line dependent (Hino et al., 2005). While we found both phosphorylation events occur in the presence of PGE2 in vivo, it remains to be determined if phosphorylation of β-catenin and GSK3β each play a significant role in the regulation of wnt activity by PGE2 in HSCs, especially given a recently proposed functional redundancy of GSK3β with GSK3α (Doble et al., 2007).

PGE2 may have co-evolved with Wnt as a mechanism to rapidly upregulate cellular proliferation to foster organ repair. In this setting, PGE2 - which is produced locally in response to tissue damage - is required for and can enhance the proliferative effects initiated by wnt activitation.

Evolutionarily, what was greatly beneficial for wound healing – the coordinated pro-proliferative, anti-apoptotic effects of wnt and PGE2, may be detrimental in cases of chronic inflammation or constitutive pathway activation and lead to tumor initiation and growth.

 

[polishkickbuttowski]

Well this sounds interesting. I noticed that Way-262611 was used in this paper whereas Way-31660 is the one commonly talked about with hair loss, is that a significant difference? Also what do you think would be an effective daily dose of gallocyanine?

 

[wislow9]

WAY-262611 is a dkk inhibitor, and WAY-316606 is an SFRP inhibitor. WAY-316606 is the one that was tested for hair loss, but it seems that WAY-262611 might be just as good, and might work synergistically.

If I try Gallocyanine I'll probably use at least 10mg once a day.

SM04554 is power dkk1 inhibitor, and we know the test dose, 0,15%.
Wouldn't SM + way316606 be better than way262611 + way316606 ¿?

 

[pegasus2]

SM04554 is power dkk1 inhibitor, and we know the test dose, 0,15%.
Wouldn't SM + way316606 be better than way262611 + way316606 ¿?

No, SM is a GSK3B inhibitor. I would prefer a Wnt agonist that inhibits dkk or sfrp. There are question marks around GSK3B inhibiton. With the paltry trial results from SM, and the failure of lithium in Follica's trial, I'm inclined to believe it could do more harm than good.

 

[John Difool]

SM trial results show hair regrowth from baseline even after stopping treatment. The improvements weren't significant but hair increase is a good sign right?

 

[pegasus2]

I guess it depends on the interplay between other pathways. Maybe it would be beneficial to inhibit GSK3B, SFRP, DKK, and Wif1 all at the same time, or maybe that would cause excessive Wnt signalling to the point that it creates cell stress and depletes the stem cell population. Minoxidil already inhibits GSK3B by upregulating AKT, which phosporylates GSK3B. It's probably a balance that we need. Are we going to inhibit it too much, and therefore increase canonical Wnt signaling too much, causing cell stress and depletion, or causing excessive mTOR activity because GSK3B inhibits mTOR. This last paper suggests that constitutive mTOR activity is the cause of cell depletion and damage rather than b-catenin itself.

Second, sustained (chronic) stimulation of either PGE2 or WNT signaling could potentially have the opposite effect and deplete stem cell activity (Kirstetter et al., 2006, Scheller et al., 2006). Therefore, it might be appropriate in a clinical setting to modulate β-catenin through these pathways, but there can be too much of a “good thing.” It will be a tricky balancing act to promote regeneration without depletion of stem cell pools or promotion of oncogenesis.
https://www.sciencedirect.com/science/article/pii/S1934590909001076

Overexpression of Wnt10b not only accelerates hair follicle to enter anagen phase, but also promotes melanocytes differentiation in young adult mice (2-month old), with increased β-catenin expression in melanocytes at the secondary hair germ and matrix region of regenerated hair follicles. Overexpression of Wnt10b also promotes melanocyte progenitor cells differentiation in vitro. Our data suggest that increased Wnt signaling promotes excessive differentiation of melanocytes, leading to exhaustion of melanocyte stem cells and eventually canities in aged mice.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5642481/

While the exhaustion of the stem cells may act as a protective mechanism against oncogenic perturbation of this particular self-renewing cell population, the persistent activation of mTOR may also contribute to accelerating aging, hence providing a novel molecular target for pharmacological intervention in multiple diseases that are characterized by the pathological depletion of stem cells, loss of tissue regenerative capacity, and tissue aging.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2939833/

 

[pegasus2]

One extreme thought is to use all the Wnt agonists together, plus rapamycin.

 

[polishkickbuttowski]

It sounds like gallocyanine is some kind of laboratory dye, so wouldn't it turn our scalps and hair purple? If it somehow doesn't I'd love to give it a shot.

 

[pegasus2]

It sounds like gallocyanine is some kind of laboratory dye, so wouldn't it turn our scalps and hair purple? If it somehow doesn't I'd love to give it a shot.

It is. I don't know if it stains skin or not.

 

[InBeforeTheCure]

I guess it depends on the interplay between other pathways. Maybe it would be beneficial to inhibit GSK3B, SFRP, DKK, and Wif1 all at the same time, or maybe that would cause excessive Wnt signalling to the point that it creates cell stress and depletes the stem cell population. Minoxidil already inhibits GSK3B by upregulating AKT, which phosporylates GSK3B. It's probably a balance that we need. Are we going to inhibit it too much, and therefore increase canonical Wnt signaling too much, causing cell stress and depletion, or causing excessive mTOR activity because GSK3B inhibits mTOR. This last paper suggests that constitutive mTOR activity is the cause of cell depletion and damage rather than b-catenin itself.

Second, sustained (chronic) stimulation of either PGE2 or WNT signaling could potentially have the opposite effect and deplete stem cell activity (Kirstetter et al., 2006, Scheller et al., 2006). Therefore, it might be appropriate in a clinical setting to modulate β-catenin through these pathways, but there can be too much of a “good thing.” It will be a tricky balancing act to promote regeneration without depletion of stem cell pools or promotion of oncogenesis.
https://www.sciencedirect.com/science/article/pii/S1934590909001076

Overexpression of Wnt10b not only accelerates hair follicle to enter anagen phase, but also promotes melanocytes differentiation in young adult mice (2-month old), with increased β-catenin expression in melanocytes at the secondary hair germ and matrix region of regenerated hair follicles. Overexpression of Wnt10b also promotes melanocyte progenitor cells differentiation in vitro. Our data suggest that increased Wnt signaling promotes excessive differentiation of melanocytes, leading to exhaustion of melanocyte stem cells and eventually canities in aged mice.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5642481/

While the exhaustion of the stem cells may act as a protective mechanism against oncogenic perturbation of this particular self-renewing cell population, the persistent activation of mTOR may also contribute to accelerating aging, hence providing a novel molecular target for pharmacological intervention in multiple diseases that are characterized by the pathological depletion of stem cells, loss of tissue regenerative capacity, and tissue aging.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2939833/

Spatial distribution of Wnt activation might be a big problem. GSK3 inhibitors activate Wnt signaling broadly, whereas cyclosporine or SFRP inhibitors (for example) amplify endogenous Wnt signaling. In that case, Wnt activation is stronger but its distribution is relatively unchanged within the HF.

 

[pegasus2]

Spatial distribution of Wnt activation might be a big problem. GSK3 inhibitors activate Wnt signaling broadly, whereas cyclosporine or SFRP inhibitors (for example) amplify endogenous Wnt signaling. In that case, Wnt activation is stronger but its distribution is relatively unchanged within the HF.

We probably only need Wnt activation in the DP, right?

 

[InBeforeTheCure]

We probably only need Wnt activation in the DP, right?

During early anagen: DP, dermal sheath cup, secondary germ, bulge
During full anagen: DP, matrix, maybe bulge?, maybe dermal sheath cup?

Keep it quiet where it's supposed to be quiet - in the dermal sheath and outer root sheath.

 

[pegasus2]

During early anagen: DP, dermal sheath cup, secondary germ, bulge
During full anagen: DP, matrix, maybe bulge?, maybe dermal sheath cup?

Keep it quiet where it's supposed to be quiet - in the dermal sheath and outer root sheath.

Seems like an impossible task. I don't suppose you have a link that shows every Wnt inhibitor's spatial expression within the hair follice? My stack now consists of WAY-316606 for SFRP, and PGE2 and estrogen covering GSK and DKK.

 

[InBeforeTheCure]

Seems like an impossible task. I don't suppose you have a link that shows every Wnt inhibitor's spatial expression within the hair follice? My stack now consists of WAY-316606 for SFRP, and PGE2 and estrogen covering GSK and DKK.

Far from impossible! The Wnt-inactive regions don't receive Wnt ligands. With WAY-316606 for example, you inhibit SFRP to increase the number of free Wnts without activating Wnt signaling in those other regions. So you can use it broadly to increase Wnt signaling only in exactly those places that have a certain level of Wnt signaling already. Look at Fig. 2 here. (LEF1 and Axin2 mRNA are readouts for Wnt activity)

 

[pegasus2]

Far from impossible! The Wnt-inactive regions don't receive Wnt ligands. With WAY-316606 for example, you inhibit SFRP to increase the number of free Wnts without activating Wnt signaling in those other regions. So you can use it broadly to increase Wnt signaling only in exactly those places that have a certain level of Wnt signaling already. Look at Fig. 2 here. (LEF1 and Axin2 mRNA are readouts for Wnt activity)

That makes sense. What do you make of Follica getting lower hair counts with the inclusion of a GSK3b inhibitor versus without it?

 

[pegasus2]

I'm going to start taking rapamycin. It is supposed to be beneficial anyways.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6814615/

Let's see if we can compile a list of Wnt agonists in this thread and what they antagonize or agonize.

LITHIUM: GSK3
SM04554: GSK3
WAY-316606: SFRP1
WAY-262611: DKK1
GALLOCYANINE: DKK1
ESTROGEN: DKK1 + GSK3
PGE2: GSK3

 

[InBeforeTheCure]

That makes sense. What do you make of Follica getting lower hair counts with the inclusion of a GSK3b inhibitor versus without it?

Link?

 

[John Difool]

I take both metformin (1000mg) and rapamycin (1g) daily with revestraol & nmn.

I also added α-KB to my list of supplements.

Rapamycin boots T so that may play a role in DHT production and T effect on HF.

https://www.sciencedirect.com/science/article/pii/S2211124719306990