Dickkopf vs TGF-Beta

[docj077]

They are not confusing to me, or anyone else who can see the obvious connections. As Michael pointed out, it is a pity there was no hair present to test as in the original study in this thread.

And we do have a clear example for ineficient lymphatics in the scalp of "HUMANS", and follicle miniaturisation. This has been studied histologicaly in lipedemous alopecia.

http://alopecia.researchtoday.net/archive/1/2/66.htm

Quote.

". CONCLUSIONS: Our findings suggest a lessened role of racial factors, but confirms the sex implications in these related conditions, and stress the potential significance of lymphangiectatic vessels in the development of alopecia in these patients."

You can theorise all you want about what particular molecules etc play in male pattern baldness, but this was a "hard" in-vivo human study that shows your assumptions above to be wrong Doctor. Of course any growth restriction in follicles for "whatever" reason, is "THEN" going to show growth related gene expression changes. But this tells us nothing about the actual "cause" of the changes related to DHT. But i get sick of trying to point out the obvious!

S Foote.

Lipedemous alopecia is a different disease and it isn't even common to men. You keep confusing them. They demonstrate different histopathologies.

As for the molecular basis of hair loss, I think that it is you who should be concerned. At the gene level, lymphedema and male pattern baldness are two very, very different processes. At the histological level, they aren't similar at all.

 

[michael barry]

Stephen,

I'll definitely point that out for Armando.....





Harold,

You are "coming up against" some of the same kinda quandries I have when considering "ideas for topicals". It seems like if something is terrifically anti-androgenic, it will inhibit VEGF or angiogenesis. If something might not inhibit angiogenesis and is theoretically anti-androgenic (beta sitosterol), I find that it ups TGF-beta expression. Something like menthol, which Ive watched literall make half my chin look like a 20 year olds with a sparse beard, probably slows keratinocyte cell division down (Alpecin's experiments) somewhat. Ive thought about topical black tea extract (high in caffeine, which might "bind" Dippkopf, inhibit TGF-beta, inhibit alpha five reductase, and downregs androgen receptors in prostate cancer cells), but hell.........................Im sure it will be shown to do something negative also. Grape seed proanthocyandins, which inhibit PKC, have been found to cause apoptosis in some cell or another. I have run into stuff like this all the time when spending time with google thinking about whatever concoction I could make at home. Ive come up with nothing that seems hunky-dory on all points. Its frustrating. Ive been hoping against hope there would be a topical out there we could easily make at home that would be anti-androgenic, anti-oxidant, pro-vegf, and inhibit tgf-beta and dkk-1.....................even if it was a combo of two or three things. Ive been unsuccessful------and hence am still on finasteride and nizoral and prox-n.

 

[joseph49853]

This is perhaps the most fascinating and illuminating discussion on the entire forum. I believe these spotlighted factors and processes -- in order of importance -- with a developmental understanding of the safety and efficacy in enhancement/suppression, not only is at the real crux of hairloss, but as well the overall inflammation, oxidation, calcification, mutations, scaring, immunosuppression, immunodeficiency, malnutrition, toxicities etc. responsible for modern diseases.

The natural and proper solutions are sitting right in front of us, but medical science is too busy looking at patentable versions alone. Meanwhile, mostly pioneering supplement companies have somewhat ameliorated the situation by filling this systematic void. The average person is looking to break free and simply doesn't know where to start. I would also like to see more comprehensively targeted, honest, practical, creative, maverick attempts at solutions, and less profit driven approaches, like housed in the below pdf... speaking of wound healing, and both the pros and cons of VEGF and angiogenesis.

A Novel Approach to Cancer Treatment: Botanicals that Inhibit Angiogenesis and/or Enhance Nonspecific Biological Immune Response

 

[harold]

a quick search on the third most upregulated gene in androgen stmulated follicles yielded this:

The Critical Roles of Serum/Glucocorticoid-Regulated Kinase 3 (SGK3) in the Hair Follicle Morphogenesis and Homeostasis - The Allelic Difference Provides Novel Insights into Hair Follicle Biology

Authors: Okada, Taro; Ishii, Yoshiyuki; Masujin, Kentaro; Yasoshima, Akira; Matsuda, Junichiro; Ogura, Atsuo; Nakayama, Hiroyuki; Kunieda, Tetsuo; Doi, Kunio

Source: American Journal of Pathology, Volume 168, Number 4, 1 April 2006 , pp. 1119-1133(15)

Abstract:
Mutation in the serum/glucocorticoid regulated kinase 3 ( Sgk3 , also known as Sgkl or Cisk ) gene causes both defective hair follicle development and altered hair cycle in mice. We examined Sgk3 -mutant YPC mice (YPC- Sgk3 ypc / Sgk3 ypc ) and found expression of SGK3 protein with altered function. In the hair follicles of YPC mice, the aberrant differentiation and poor proliferation of hair matrix keratinocytes during the period of postnatal hair follicle development resulted in a complete lack of hair medulla and weak hair. Surprisingly, the length of postnatal hair follicle development and anagen term was shown to be dramatically shortened. Also, phosphorylation of GSK3? at Ser9 and the nuclear accumulation of ?-catenin were reduced in the developing YPC hair follicle, suggesting that phosphorylation of GSK3? and WNT-?-catenin pathway takes part in the SGK3-dependent regulation of hair follicle development. Moreover, the above-mentioned features, especially the hair-cycling pattern, differ from those in other Sgk3 -null mutant strains, suggesting that the various patterns of dysfunction in the SGK3 protein may result in phenotypic variation. Our results indicate that SGK3 is a very important and characteristic molecule that plays a critical role in both hair follicle morphogenesis and hair cycling.

Document Type: Research article


Looks like this protein also interacts with the wnt pathway. Will try to read more later.
hh

 

[harold]

More to dump in this thread
So TgF-Beta1/2 are important in promoting catagen. It looks lik completely abrogating their function actually leads to hair loss in the long term as hair follicles were disturbed from their normal cycle of growth and regression. As with some of the stuff showing that while acute signalling of the wnt pathway leads to hair formation while chronic signalling leads to aberrant hair growth/tumorgenesis it looks like too much of a good thing is a bad thing. Or in this case - not enough of a bad thing is a bad thing.


Cancer Res. 2005 Oct 1;65(19):8671-8.Click here to read Links
Targeted disruption of Smad4 in mouse epidermis results in failure of hair follicle cycling and formation of skin tumors.
Yang L, Mao C, Teng Y, Li W, Zhang J, Cheng X, Li X, Han X, Xia Z, Deng H, Yang X.

Genetic Laboratory of Development and Diseases, Institute of Biotechnology, Shanghai, P.R.China.

Smad4 is the common mediator of transforming growth factor-beta (TGF-beta) superfamily signaling, which functions in diverse developmental processes in mammals. To study the role of Smad4 in skin development, a keratinocyte-specific null mutant of Smad4 (Smad4(co/co);K5-Cre) was generated in mice using the Cre-loxP system. The Smad4-mutant mice exhibited progressive alopecia as a result of the mutant hair follicles failing to undergo programmed regression. Sonic hedgehog (Shh) was only detected in Smad4-mutant hair follicles at the catagen stage. Seventy percent of Smad4(co/co); K5-Cre mice developed spontaneous tumors within 12 months of birth. c-Myc and cyclin D1 were up-regulated whereas p21 and p27 expressions were decreased, which correlated with the epidermal hyperplasia in Smad4 mutants. Interestingly, coordinated deletion of the Smad4 and PTEN genes resulted in accelerated hair loss and skin tumor formation, suggesting that Smad4 and PTEN act synergistically to regulate epidermal proliferation and differentiation. All of our data indicate that Smad4 is essential for catagen induction and acts as a critical suppressor in skin tumorigenesis.

 

[harold]

Here is some more on the effect of chronic stimulation of the wnt/beta-canenin pathway vs acute stimulation:

Genes Dev. 2003 May 15;17(10):1219-24.Click here to read Links
Transient activation of beta -catenin signaling in cutaneous keratinocytes is sufficient to trigger the active growth phase of the hair cycle in mice.
Van Mater D, Kolligs FT, Dlugosz AA, Fearon ER.

Departments of Human Genetics, University of Michigan School of Medicine, Ann Arbor, Michigan 48109, USA.

Wnts have key roles in many developmental processes, including hair follicle growth and differentiation. Stabilization of beta-catenin is essential in the canonical Wnt signaling pathway. We developed transgenic mice expressing a regulated form of beta-catenin in the skin. Chronic activation of beta-catenin in resting (telogen) hair follicles resulted in changes consistent with induction of an exaggerated, aberrant growth phase (anagen). Transient activation of beta-catenin produced a normal anagen. Our data lend strong support to the notion that a Wnt/beta-catenin signal operating on hair follicle precursor cells serves as a crucial proximal signal for the telogen-anagen transition.

and more....


Development. 2004 Apr;131(8):1787-99.Click here to read Links
Transient activation of beta-catenin signalling in adult mouse epidermis is sufficient to induce new hair follicles but continuous activation is required to maintain hair follicle tumours.
Lo Celso C, Prowse DM, Watt FM.

Keratinocyte Laboratory, Cancer Research UK London Research Institute, 44 Lincoln's Inn Fields, London WC2A 3PX, UK.

When beta-catenin signalling is disturbed from mid-gestation onwards lineage commitment is profoundly altered in postnatal mouse epidermis. We have investigated whether adult epidermis has the capacity for beta-catenin-induced lineage conversion without prior embryonic priming. We fused N-terminally truncated, stabilised beta-catenin to the ligand-binding domain of a mutant oestrogen receptor (DeltaNbeta-cateninER). DeltaNbeta-cateninER was expressed in the epidermis of transgenic mice under the control of the keratin 14 promoter and beta-catenin activity was induced in adult epidermis by topical application of 4-hydroxytamoxifen (4OHT). Within 7 days of daily 4OHT treatment resting hair follicles were recruited into the hair growth cycle and epithelial outgrowths formed from existing hair follicles and from interfollicular epidermis. The outgrowths expressed Sonic hedgehog, Patched and markers of hair follicle differentiation, indicative of de novo follicle formation. The interfollicular epidermal differentiation program was largely unaffected but after an initial wave of sebaceous gland duplication sebocyte differentiation was inhibited. A single application of 4OHT was as effective as repeated doses in inducing new follicles and growth of existing follicles. Treatment of epidermis with 4OHT for 21 days resulted in conversion of hair follicles to benign tumours resembling trichofolliculomas. The tumours were dependent on continuous activation of beta-catenin and by 28 days after removal of the drug they had largely regressed. We conclude that interfollicular epidermis and sebaceous glands retain the ability to be reprogrammed in adult life and that continuous beta-catenin signalling is required to maintain hair follicle
tumours.

Not sure quite what to make of this one - reports that sutained high level activation can form new follicles out of old ones while low level converts sebaceous glands and skin cells into follicles.

Dev Cell. 2005 Jul;9(1):121-31.Click here to read Links
Beta-catenin and Hedgehog signal strength can specify number and location of hair follicles in adult epidermis without recruitment of bulge stem cells.
Silva-Vargas V, Lo Celso C, Giangreco A, Ofstad T, Prowse DM, Braun KM, Watt FM.

Keratinocyte Laboratory, Cancer Research UK London Research Institute, 44 Lincoln's Inn Fields, London WC2A 3PX, United Kingdom.

Using K14deltaNbeta-cateninER transgenic mice, we show that short-term, low-level beta-catenin activation stimulates de novo hair follicle formation from sebaceous glands and interfollicular epidermis, while only sustained, high-level activation induces new follicles from preexisting follicles. The Hedgehog pathway is upregulated by beta-catenin activation, and inhibition of Hedgehog signaling converts the low beta-catenin phenotype to wild-type epidermis and the high phenotype to low. beta-catenin-induced follicles contain clonogenic keratinocytes that express bulge markers; the follicles induce dermal papillae and provide a niche for melanocytes, and they undergo 4OHT-dependent cycles of growth and regression. New follicles induced in interfollicular epidermis are derived from that cellular compartment and not through bulge stem cell migration or division. These results demonstrate the remarkable capacity of adult epidermis to be reprogrammed by titrating beta-catenin and Hedgehog signal strength and establish that cells from interfollicular epidermis can acquire certain characteristics of bulge stem cells.

 

[harold]

OK so the next most upregulated gene after dickkopf - SGK can act as a GSK3 inhibitor. Inhibiting GSK3 leads to the accumulation of Beta-Catenin and thus enhanced wnt signalling. Seriously wtf? Why couldnt it have worked the EXACT OPPOSITE way? That would make sense at least.....
hh

Mol Endocrinol. 2007 Oct;21(10):2403-15. Epub 2007 Jun 26.Click here to read Links
Glucocorticoid-induced degradation of glycogen synthase kinase-3 protein is triggered by serum- and glucocorticoid-induced protein kinase and Akt signaling and controls beta-catenin dynamics and tight junction formation in mammary epithelial tumor cells.
Failor KL, Desyatnikov Y, Finger LA, Firestone GL.

Department of Molecular and Cell Biology, University of California at Berkeley, Berkeley, California 94720-3200, USA.

Glucocorticoid hormones stimulate adherens junction and tight junction formation in Con8 mammary epithelial tumor cells and induce the production of a stable nonphosphorylated beta-catenin protein localized exclusively to the cell periphery. Glycogen synthase kinase-3 (GSK3) phosphorylation of beta-catenin is known to trigger the degradation of this adherens junction protein, suggesting that steroid-activated cascades may be targeting this protein kinase. We now demonstrate that treatment with the synthetic glucocorticoid dexamethasone induces the ubiquitin-26S proteasome-mediated degradation of GSK3 protein with no change in GSK3 transcript levels. In transfected cells, deletion of the N-terminal nine amino acids or mutation of the serine-9 phosphorylation site on GSK3-beta prevented its glucocorticoid-induced degradation. Expression of stabilized GSK3 mutant proteins ablated the glucocorticoid-induced tight junction sealing and resulted in production of a nonphosphorylated beta-catenin that localizes to both the nucleus and the cell periphery in steroid-treated cells. Serine-9 on GSK3 can be phosphorylated by Sgk (serum- and glucocorticoid-induced protein kinase) and by Akt. Expression of dominant-negative forms of either Sgk- or Akt-inhibited glucocorticoid induced GSK3 ubiquitination and degradation and disrupted the dexamethasone-induced effects on beta-catenin dynamics. Furthermore, the steroid-induced tight junction sealing is attenuated in cells expressing dominant-negative forms of either Sgk or Akt, although the effect of blunting Sgk signaling was significantly greater. Taken together, we have uncovered a new cellular cascade in which Sgk and Akt trigger the glucocorticoid-regulated phosphorylation, ubiquitination, and degradation of GSK3, which alters beta-catenin dynamics, leading to the formation of adherens junctions and tight junction sealing.

 

[harold]

OK so it is SGK3 that is important for hair ssurvival by the pathway suggested above it seems -it functions as a GSK3 inhibitor like lithium.

Am J Pathol. 2006 Apr;168(4):1119-33.Click here to read Click here to read Links
The critical roles of serum/glucocorticoid-regulated kinase 3 (SGK3) in the hair follicle morphogenesis and homeostasis: the allelic difference provides novel insights into hair follicle biology.
Okada T, Ishii Y, Masujin K, Yasoshima A, Matsuda J, Ogura A, Nakayama H, Kunieda T, Doi K.

Department of Veterinary Pathology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-8657, Japan. aa37158@mail.ecc.u-tokyo.ac.jp

Mutation in the serum/glucocorticoid regulated kinase 3 (Sgk3, also known as Sgkl or Cisk) gene causes both defective hair follicle development and altered hair cycle in mice. We examined Sgk3-mutant YPC mice (YPC-Sgk3(ypc)/Sgk3(ypc)) and found expression of SGK3 protein with altered function. In the hair follicles of YPC mice, the aberrant differentiation and poor proliferation of hair matrix keratinocytes during the period of postnatal hair follicle development resulted in a complete lack of hair medulla and weak hair. Surprisingly, the length of postnatal hair follicle development and anagen term was shown to be dramatically shortened. Also, phosphorylation of GSK3beta at Ser9 and the nuclear accumulation of beta-catenin were reduced in the developing YPC hair follicle, suggesting that phosphorylation of GSK3beta and WNT-beta-catenin pathway takes part in the SGK3-dependent regulation of hair follicle development. Moreover, the above-mentioned features, especially the hair-cycling pattern, differ from those in other Sgk3-null mutant strains, suggesting that the various patterns of dysfunction in the SGK3 protein may result in phenotypic variation. Our results indicate that SGK3 is a very important and characteristic molecule that plays a critical role in both hair follicle morphogenesis and hair cycling.

It also appears that mice lacking SGK3 have the hair abnormalities but mice lacking SGK/SGK1 do not. So SGK does not seem to perform the same GSK3 inhibition.

Mol Biol Cell. 2004 Sep;15(9):4278-88. Epub 2004 Jul 7.
Targeted disruption of the protein kinase SGK3/CISK impairs postnatal hair follicle development.
McCormick JA, Feng Y, Dawson K, Behne MJ, Yu B, Wang J, Wyatt AW, Henke G, Grahammer F, Mauro TM, Lang F, Pearce D.

Departments of Medicine and Molecular and Cellular Pharmacology, University of California, San Francisco, CA 94143, USA.

Members of the serum- and glucocorticoid-regulated kinase (SGK) family are important mediators of growth factor and hormone signaling that, like their close relatives in the Akt family, are regulated by lipid products of phosphatidylinositol-3-kinase. SGK3 has been implicated in the control of cell survival and regulation of ion channel activity in cultured cells. To begin to dissect the in vivo functions of SGK3, we generated and characterized Sgk3 null mice. These mice are viable and fertile, and in contrast to mice lacking SGK1 or Akt2, respectively, display normal sodium handling and glucose tolerance. However, although normal at birth, by postpartum day 4 they have begun to display an unexpected defect in hair follicle morphogenesis. The abnormality in hair follicle development is preceded by a defect in proliferation and nuclear accumulation of beta-catenin in hair bulb keratinocytes. Furthermore, in cultured keratinocytes, heterologous expression of SGK3 potently modulates activation of beta-catenin/Lef-1-mediated gene transcription. These data establish a role for SGK3 in normal postnatal hair follicle development, possibly involving effects on beta-catenin/Lef-1-mediated gene transcription.

Why then might its upregulation be so high here? Perhaps the presence of high levels of SGK1 actually interfere with the performance of SGK3 or causes its downregulation?
hh

 

[Bryan]

Anyway. It seems that Dickkopf is way ahead as the number 2 most upregulated gene while TGF-Beta-2 comes in at number 30 with about half as much extra RNA produced after DHT does its thing.

You mean DHT at normal physiological concentrations, as opposed to the 100 nM used in this study? At 100 nM, TGF-Beta-2 was almost exactly doubled, it didn't just increase by half.

 

[Bryan]

The top 30 upregulated genes in balding dermal papilla cells in response to 100 nM DHT determined by microarray hybridization. <snip list of upregulated genes>

I'm shocked that nobody else has even mentioned something that seems screamingly obvious to me, which is that such a list of upregulated genes in BALDING HAIR FOLLICLES is only half the picture. It needs to be compared and contrasted with a list of upregulated genes in NON-BALDING HAIR FOLLICLES (like maybe beard follicles)!

It's not out of the question that certain genes could be upregulated by androgens in both types of hair follicles, and comparing the two lists could go a long way toward eliminating a lot of wild-goose chases by showing us which gene-product-inhibiting substances NOT to pursue. Obsessing over just the single list above seems very unwise to me.

 

[Bryan]

Michael.

This is a study you may want to point out to Armando? As i said, i don't have time anymore to get involved in arguments on these boards.

Oh my goodness...is it really because you don't have time to get involved in arguments anymore, or is it because your eccentric theory gets shot down in flames and explodes on impact every time you bring it up around here?

 

[harold]

Anyway. It seems that Dickkopf is way ahead as the number 2 most upregulated gene while TGF-Beta-2 comes in at number 30 with about half as much extra RNA produced after DHT does its thing.

You mean DHT at normal physiological concentrations, as opposed to the 100 nM used in this study? At 100 nM, TGF-Beta-2 was almost exactly doubled, it didn't just increase by half.

Looking back it was about "half as much fold increase in RNA production" which is what i should have said. I guess "half as upregulated" is less unwieldy.

Would 100nM be a hugely supraphysiological dose?
hh

 

[harold]

The top 30 upregulated genes in balding dermal papilla cells in response to 100 nM DHT determined by microarray hybridization. <snip list of upregulated genes>

I'm shocked that nobody else has even mentioned something that seems screamingly obvious to me, which is that such a list of upregulated genes in BALDING HAIR FOLLICLES is only half the picture. It needs to be compared and contrasted with a list of upregulated genes in NON-BALDING HAIR FOLLICLES (like maybe beard follicles)!

It's not out of the question that certain genes could be upregulated by androgens in both types of hair follicles, and comparing the two lists could go a long way toward eliminating a lot of wild-goose chases by showing us which gene-product-inhibiting substances NOT to pursue. Obsessing over just the single list above seems very unwise to me.

This is true. However they did go onto show that DKK-1 inhibited outer root sheath cells in vitro so it does not look like it is an incidental player by any means. The comparison between balding and non-balding hair was done in the study below way back in 2004. Again the answers are more or less completely different.

J Dermatol Sci. 2004 Oct;36(1):25-32.Click here to read Links
Different gene expression profile observed in dermal papilla cells related to androgenic alopecia by DNA macroarray analysis.
Midorikawa T, Chikazawa T, Yoshino T, Takada K, Arase S.

Biological Science Research Center, Lion Corporation, Odawara City, 100 Tajima Odawara, Kanagawa 256-0811, Japan.

BACKGROUND: Androgenic alopecia (Androgenetic Alopecia) is the most common type of baldness in men. Although etiological studies have proved that androgen is one of the causes of this symptom, the defined molecular mechanism underlying androgen-related actions remains largely unknown. OBJECTIVES: To clarify the difference in the gene expression profile of dermal papilla cells (DPCs) in skin affected by baldness. METHODS: DNA macroarray study was carried out on cultured DPCs from Androgenetic Alopecia skin comparing with DPCs from skin that is not affected by baldness. RESULTS: From DNA macroarray analysis, we observed that 107 of the 1185 analyzed genes had differing expression levels. A marked difference was observed in the decreased gene expression of BMP2 and ephrin A3 and up-regulated in NT-4 gene. In order to clarify the roles of BMP2 and ephrin A3 in the hair follicles, we examined the proliferation of hair follicle keratinocyte and expression of a hair acidic keratin gene. Both BMP2 and ephrin A3 raised the proliferation rate of the outer root sheath cells (ORSCs) and induced gene expression in acidic hair keratin 3-II. Conclusion: These results lead us to the hypothesis that both BMP2 and ephrin A3 function as hair growth promoting factors in the hair cycle.

Again in Cotsarelis's newest patent they claim to have done this and come up with a huge difference in the expression of PGD2 in balding and non-balding regions and to a lesser extent PGE2 (more of this in non-balding).....some consistency would be nice. I guess it also helps to look what else we know about proteins like DKK-1 and PGD2 and try to work out what role they might play - one may be clearly more important.
hh

 

[Bryan]

Anyway. It seems that Dickkopf is way ahead as the number 2 most upregulated gene while TGF-Beta-2 comes in at number 30 with about half as much extra RNA produced after DHT does its thing.

You mean DHT at normal physiological concentrations, as opposed to the 100 nM used in this study? At 100 nM, TGF-Beta-2 was almost exactly doubled, it didn't just increase by half.

Looking back it was about "half as much fold increase in RNA production" which is what i should have said. I guess "half as upregulated" is less unwieldy.

You've lost me. Where did it say that it was "half as much fold increase..."?

Would 100nM be a hugely supraphysiological dose?

It would be just for blood levels, yes. Checking the Merck Manual for average serum levels of steroid hormones, it says the average for testosterone in an adult male is from 10.4 - 38.1 nM/L. It doesn't have an entry for DHT, but I know from other sources that serum DHT is approximately 1/10 that of serum testosterone, implying that the average for DHT would be something roughly on the order of 1 - 4 nM/L. That's a FAR cry from 100 nM!

Edit: But the problem here is that I don't really know for sure what the normal physiological levels of DHT for tissue levels like dermal papillae would be. My hunch is that it would be more in line with testosterone levels, which would still mean that 100 nM is above normal.

 

[harold]

"Anyway. It seems that Dickkopf is way ahead as the number 2 most upregulated gene while TGF-Beta-2 comes in at number 30 with about half as much extra RNA produced after DHT does its thing."

You mean DHT at normal physiological concentrations, as opposed to the 100 nM used in this study? At 100 nM, TGF-Beta-2 was almost exactly doubled, it didn't just increase by half.

Looking back it was about "half as much fold increase in RNA production" which is what i should have said. I guess "half as upregulated" is less unwieldy.

You've lost me. Where did it say that it was "half as much fold increase..."?

Just that the numbers given were the fold increase in RNA production. So as you pointed out TGF-B2 increased about 2 fold. And DKK-1 increased almost 5 fold. Which is more or less twice the "fold increase" of TGF-B2......it was an awkward thing to say and i couldnt tell what i was on about myself when you quoted it. But I think thats what I must have meant.

Would 100nM be a hugely supraphysiological dose?

It would be just for blood levels, yes. Checking the Merck Manual for average serum levels of steroid hormones, it says the average for testosterone in an adult male is from 10.4 - 38.1 nM/L. It doesn't have an entry for DHT, but I know from other sources that serum DHT is approximately 1/10 that of serum testosterone, implying that the average for DHT would be something roughly on the order of 1 - 4 nM/L. That's a FAR cry from 100 nM!

Edit: But the problem here is that I don't really know for sure what the normal physiological levels of DHT for tissue levels like dermal papillae would be. My hunch is that it would be more in line with testosterone levels, which would still mean that 100 nM is above normal.

Yeah it does seem like it must be a lot more. Or "several fold" more! Maybe not.
While I guess it doesnt reflect what is happening exactly in the follicle I would rather they "overdid it" than the opposite. I just wish there was more accord between these studies. Someone has to come along and look at all thesee things at once - DKK-1 PGD2 TGF-B and you are right there needs to be non-balding results to compare with.
hh

 

[harold]

The top 30 upregulated genes in balding dermal papilla cells in response to 100 nM DHT determined by microarray hybridization. <snip list of upregulated genes>

I'm shocked that nobody else has even mentioned something that seems screamingly obvious to me, which is that such a list of upregulated genes in BALDING HAIR FOLLICLES is only half the picture. It needs to be compared and contrasted with a list of upregulated genes in NON-BALDING HAIR FOLLICLES (like maybe beard follicles)!

It's not out of the question that certain genes could be upregulated by androgens in both types of hair follicles, and comparing the two lists could go a long way toward eliminating a lot of wild-goose chases by showing us which gene-product-inhibiting substances NOT to pursue. Obsessing over just the single list above seems very unwise to me.

Just remebered as was looking through this study again today - the authors did go on to show that DKKF1 was upregulated in balding scalp as compared to non-balding scalp specifically. So they did look at this - just not for every gene in their list. And it does seem to confirm DKKF1 = bad.
hh

 

[docj077]

The top 30 upregulated genes in balding dermal papilla cells in response to 100 nM DHT determined by microarray hybridization. <snip list of upregulated genes>

I'm shocked that nobody else has even mentioned something that seems screamingly obvious to me, which is that such a list of upregulated genes in BALDING HAIR FOLLICLES is only half the picture. It needs to be compared and contrasted with a list of upregulated genes in NON-BALDING HAIR FOLLICLES (like maybe beard follicles)!

It's not out of the question that certain genes could be upregulated by androgens in both types of hair follicles, and comparing the two lists could go a long way toward eliminating a lot of wild-goose chases by showing us which gene-product-inhibiting substances NOT to pursue. Obsessing over just the single list above seems very unwise to me.

Just remebered as was looking through this study again today - the authors did go on to show that DKKF1 was upregulated in balding scalp as compared to non-balding scalp specifically. So they did look at this - just not for every gene in their list. And it does seem to confirm DKKF1 = bad.
hh

DKKF1 is not necessarily bad. Sure, it's harmful for hair growth and proper cycling, but it must have a reason for being upregulated. The same can be said for TGF-beta. The human body doesn't just produce negative regulators of proliferative pathways, because it feels like it. There has to be a signal that is either hyperproliferative or simply overabundant and genetically degenerative in order for such changes to take place.

My guess is that even knowing the molecular basis of hairloss in terms of which negative regulators are upregulated will prove rather useless to us until we know why they are upregulated. And, simply saying that it's because of increased androgen response is not enough. There has to be a reason for that, as well. Find that, and you'll have your cure.

Messing with cellular signaling pathways that are as tightly controlled as the Wnt signaling pathway (ie inhibiting DKK1 function) is fruitless and naive on the part of those that even attempting it. The potential damage that such pharmacological or procedural inventions could have on the human body will be both deadly and unforgivable.

 

[harold]

DKKF1 is not necessarily bad. Sure, it's harmful for hair growth and proper cycling,

True its not bad but its "bad" like dht is "bad" - as you say not good for promoting scalp hair. Unlike DHT though DKKF1 is more or less bad news for any hair.

but it must have a reason for being upregulated. The same can be said for TGF-beta. The human body doesn't just produce negative regulators of proliferative pathways, because it feels like it. There has to be a signal that is either hyperproliferative or simply overabundant and genetically degenerative in order for such changes to take place.

This is where I begin to disagree with you - I dont think there has to be any reason at all for DHT to produce DKKF1. I dont think there is any particular reason to have hair on the top of your head either apart from cosmetic/interpersonal ones. Millions of people get by fine every day without it. I dont think there has to be a hyperproliferative signal to trigger the release of DKKF1 (though antiproliferative signals obviously protect against tumour development.)

My guess is that even knowing the molecular basis of hairloss in terms of which negative regulators are upregulated will prove rather useless to us until we know why they are upregulated. And, simply saying that it's because of increased androgen response is not enough. There has to be a reason for that, as well. Find that, and you'll have your cure.

I dont think there has to be a "why". The body doesn't always have a reason for everything it does - things just go wrong. Its amazing that they dont go more wrong more often. If this is not what you are saying then I apologise - when I first started posting here people thought I was some vitamin freak cos of my interest in ROS signalling in male pattern baldness as you probably remember. But yeah I think it could and probably is as simple as androgens -> negative regulators. Though an increased inflammatory response and a loss of "immune privilege" by the hair follicles probably has a big role somewhere in the middle. But I dont see that as a "why" myself - just another link in a chain of events.

Messing with cellular signaling pathways that are as tightly controlled as the Wnt signaling pathway (ie inhibiting DKK1 function) is fruitless and naive on the part of those that even attempting it. The potential damage that such pharmacological or procedural inventions could have on the human body will be both deadly and unforgivable.

I disagree - done in a localised and controlled manner I dont see the problem. Research seems to indicate that androgens are messing with such tightly controlled signalling pathways on both of our heads even as we speak.
hh

 

[docj077]

Messing with cellular signaling pathways that are as tightly controlled as the Wnt signaling pathway (ie inhibiting DKK1 function) is fruitless and naive on the part of those that even attempting it. The potential damage that such pharmacological or procedural inventions could have on the human body will be both deadly and unforgivable.

I disagree - done in a localised and controlled manner I dont see the problem. Research seems to indicate that androgens are messing with such tightly controlled signalling pathways on both of our heads even as we speak.
hh

You don't see the problem, because you don't understand the significance. You do not have to maintain DKK1 inhibition for long periods of time in order to disregulate the system. All it takes the removal of such an inhibitory signal for even a short amount of time to kick even one cell into a pro-proliferative state. This bullcrap that everyone doing this research is saying about the process being short-lived, intermittent, etc. is going to come back to bite them all in the ***. The reason this worksd for androgens for prolonged periods is because they maintain the tightly controlled system in the form of negative regulators.

Also, simply upregulating Wnt signaling will likely not regrow hair in everyone. Case and point is the recent study out of Stanford that demonstrated that the Wnt signaling pathway can have a negative effect on stem cell function. According to their research, inhibition of the pathway may be a therapeutic target to help slow aging and increase healing.

 

[michael barry]

Doctor,

in some embodiments in that patent abrasion is performed and 11 days later minoxidil is administered daily until hair germ detection AND THAT IS IT. New hair is detected later on.


Are you going to claim that dermabrasion is "dangerous" or something? Minoxdil is taken internally for other things, so it cant be "dangerous".


Ive looked into this deeper, and we probably wont even need to inhibit wnt with DKK-1 for the first nine days because human beings have pigmented skin (and could easily get some sun on their heads a week or two before the procedure to increase skin pigmentation). They admit this much in the interview.



Simply stated, they abrade the skin with a felt-wheeled instrument that results in "shiny" skin with no blood.
They may OR MAY NOT ask you to take a EGF-receptor blocker for nine days (or apply it topically)
They may OR MAY NOT ask you to put on some topical FGF at day 11 onwards to about day 20
They may OR MAY NOT ask you to use minoxidil on day 11 onwards to about day 20.


In one experiment (the first one) they noted hair growth when the mice were abraded and the wound simply was not closed or bandaged or had any ointment applied to it. Thats it.......................just abrasion.


Here is one experiment where they grew hair by abrasion and minoxidil ONLY:

EXAMPLE 16

ENHANCEMENT OF EDIHN BY ADMINISTRATION OF MINOXIDIL

[000235] To determine the effect of minoxidil on EDIHN, recombinant FGF is administered 11 days after incisional wounding, as decribed in Example 11. Minoxidil administration enhances HF formation, showing that new HF can be generated by (a) disrupting the epidermis; and (b) administering a minoxidil.





Ive read the bios of the people researching this. One guy used to head Merk, another used to head Upjohn. George Costariailis and Kurt Stenn were involved with this. No animals have gotten any tumors as of yet. By stating that you are confident that this will all lead to cancer, you are stating that ALL THESE people are wrong, have overlooked the obvious, and basically dont know what they are doing. Sorry, but Kurt Stenn probably knows more about hair and skin than anyone on the face of this planet. He is known as "the godfather of hair" at Harvard. I really, really, really, really doubt they'd pursue an indication that they thought there would be a chance of even 1% of the recipients getting cancerous tumors, dont you?




Here is another experiment whereby they grew hair by JUST PLUCKING AND THEN ABRASION ALONE:
EXPERIMENTAL DETAILS SECTION

EXAMPLE 1

DEPILATION AND EPIDERMAL ABRASION CAUSES DE NOVO HAIR FOLLICLE

FORMATION

MATERIALS AND EXPERIMENTAL METHODS

Depilation and epidermal abrasion

P-7628-PC

[000187] Mice were anesthetized with an injection of sodium pentobarbital before the hair on the back was clipped and depilated withNair (Carter-Wallace, New York, NY), then epidermis was removed using a rotating felt wheel as described by Argyris T, J Invest Dermatol, 75: 360-362, 1980). After scrubbing with 70% ethanol and drying under an incandescent lamp, the basal and supra-basal layers in an area of (1.5 cm)2 cm of the inter-folHcular epidermis were removed by careful abrasion with a felt wheel mounted on a Dremel Moto-tool (Racine, WI). After abrasion, the skin was shiny and smooth, and there was no blood. One day later, the abraded area was covered by a fibrin crust, which fell off after 3-7 days, exposing the newly regenerated epidermis. A group of control mice was sacrificed immediately after abrasion to confirm microscopically the complete removal of the interfollicular epidermis.

Immunohistochemistry

[000188] Skin samples were fixed in PBS-buffered 10% formalin. Six-micron thick paraffin sections were cut and stained, where applicable, with antibodies.

BrdU labeling [000189] The protocol described by Bickenbach and colleagues (Bickenbach et al, Cell Tiss Kinet 19: 325-333, 1986; Bickenbach et al, Exp Cell Res 244. 184-195, 1998) was used. Mice were injected with 50 milligrams per kilogram (mg/kg) bodyweight 5-bromo-2'-deoxyuridine (BrdU) every 12 hours for a total of four injections.

RESULTS

[000190] An area of the backs of 50-day old mice was subjected to depilation and removal of the epidermis using a rotating felt wheel. Fifteen days later, HF placodes, hair germs and other signs of follicle neogenesis were present (Figure 1 ; arrow indicates a hair germ). Morphology of the follicles was similar to embryonic follicle development. To further characterize proliferation in the new follicles, the skin was labeled with BrdU 60 minutes before sacrifice. As depicted in Figure 2, the proliferation pattern was similar to developing follicles in the embryo.

[000191] These findings demonstrate that (a) disruption of the epidermis causes generation of new

P-7628-PC

HF, and that this generation of new HF can occur (b) in adult subjects and (c) during telogen (50- day-old mice are in the second telogen stage of the hair cycle).