I will start Exosome treatment soon - Day 0 :-)

Arthurandsam

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Currently on Fluridil + CosmeRNA topical treatment since May 2023 (6 months) to combat the action of DHT and on 5% topical minoxidil, I am starting out with exosomes hoping to gain a little more density.

The combo I am currently using {Fluridil + CosmeRNA + Topical minoxidil} is working great on me for hair loss stabilization but I would like to have some regrow.

Here is a summary of what I have read about this area. My aim was to understand what exosomes are and how maybe they can help me in the fight against alopecia. I wanted to get an idea of the effectiveness that the ASCE+ HRLV product (which I chose to use) can have, and to try to understand how it can improve my sad condition...

You will see that after conducting research to fully understand the role that exosomes can have on alopecia, I am unfortunately a bit divided on the ASCE+ HRLV product in terms of the theoretical effectiveness that it could have. But I'm starting out anyway because there seems to be good feedback from dermatologists who use it in mesotherapy... And then alopecia is a very complex process. As I am not a doctor, I would also like to seek the help of the members of this forum to contradict, or on the contrary confirm, my ideas and remarks. Also, if I have made mistakes please let me know!

The routine that I plan to follow with these exosomes will be as follows :

  • Application in mesotherapy using 1mm multi-needles (this is the depth recommended to me by the direct representative of this product in Luxembourg)
  • Followed by a 0.5mm derma roller (this is what the researcher who intervenes in the video on this product recommends)
  • I'm going to do 5 applications spaced 2 weeks apart at the beginning
  • Then, switch to 1 application every 3 months
  • Or maybe do 1 application per month for a few months before switching to applications every 3 months.
Summary :

  1. What are exosomes ?
  2. Exosomes extracted from mesenchymal stem cells: MSC exosomes
  3. The origin of MSC exosomes plays a role in the quality of the therapeutic effect
  4. The hair cycle
  5. Hair anatomy
  6. Embryo follicle development
  7. The follicle cycle
  8. Experiments have shown that MSC-ASC exosomes cause hair growth
  9. The proteins contained in the exosomes of the ASCE+ HRLV product
  10. Alopecia and DHT
  11. The importance of the Wnt/beta-catenin signaling pathway on hair growth
  12. The canonical Wnt signaling pathway and its deterioration in alopecia
  13. DHT level is correlated with DKK1
  14. Some molecules inhibit the DKK1 protein
  15. The role of microRNAs in modulating hair growth
  16. What microRNAs are contained in ASCE+ HRLV exosomes?
  17. Conclusion
Sources :

Happy reading!

PS: I am French and therefore I wrote this text in French. This is a translation.
 
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Arthurandsam

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What are exosomes ?

The discovery of extracellular vesicles, or exosomes, dates back to the 1940s. Researchers began to focus on them only in the mid-2000s when they realized that they allow information to be exchanged between cells. There were more than 3,000 publications on this subject in 2018 and 2019... the race to commercialize therapies based on the use of exosomes is therefore on. The four main start-ups in this field are Codiak Biosciences, Exosome Diagnostics, Evox Therapeutics, and ExocoBio. They have received over 385 million dollars in investments. There have also been numerous agreements that have been created between start-ups and major pharmaceutical companies.

Exosomes are a class of extracellular vesicles (EVs — Extracellular Vesicles) of nanometric size (30nm to 200nm), released by virtually all eukaryotic cells (cells that have a nucleus bound to a membrane - the cells of our body). Exosomes can be extracted from stem cells and can have numerous therapeutic applications. These therapeutic effects result from a particular form of information transmission between cells: paracrine signaling. Indeed, these signals emitted by cells induce changes in nearby cells, which alters their behavior. These signals are transmitted at a short distance, in contrast to the signals transmitted by the endocrine system with hormones. Researchers have shown that exosomes play a very important role in the effects related to paracrine signaling.

Once the exosomes have been extracted from the stem cells and purified, they are « loaded with a cargo. » This cargo can then be transferred to other cells and thus change their cellular behavior. It contains the following materials:

  • Lipids,
  • Proteins,
  • Nucleic acids like messenger RNAs (mRNAs), microRNAs (miRNAs), and other non-coding RNAs (ncRNAs),
Lipids play the role of storing the energy that the cells of our body need to live.

Proteins are macronutrients that are the basis of every living organism. They are synthesized in the cell at the ribosome (which is located outside the nucleus), from information coded in genes (DNA is stored in the nucleus) which determines the order in which amino acids will be sequenced. Post-transcriptional modifications may occur once the protein has been synthesized, which may have the effect of altering its physical or chemical properties.

Numerous proteins, resulting from the expression of specific genes, are involved in the regulation of physiological events in the skin and hair. We will come back to this point when we study how hair is formed.

Messenger RNAs are copies of a region of DNA that are stored inside the cell nucleus. Since the ribosomes synthesizing proteins are outside the cell nucleus, when a cell needs a protein, the messenger RNA is exported outside the nucleus and joins the ribosomes where it allows the synthesis of the requested protein to be synthesized.

The rate at which genetic information is transcribed from DNA to messenger RNA is controlled by proteins called transcription factors. Their role is to activate or deactivate genes so that they are expressed at the right time and at the right level in the desired cells of the body. These transcription factors therefore have an important role in cell division, cell growth and cell death. There are 1500-1600 transcription factors in the human genome. These transcription factors can work alone or accompanied by proteins.

Non-coding RNAs correspond to all RNAs resulting from DNA transcription but which will not be translated into proteins by the ribosomal machinery. However, they are involved at numerous scales in the regulation of protein synthesis and/or the maturation of other RNAs. Among them, we find microRNAs (miRNAs), among others.

MicroRNAs are therefore non-coding RNAs that control gene expression at the post-transcriptional level. MicroRNAs regulate gene expression by pairing with target messenger RNAs with which they are partially complementary. They then inhibit the expression of their target genes, causing the blocking of protein translation or their degradation.

Numerous miRNAs have been identified as being involved in the regulation of physiological events in the skin and hair, such as the regulation of keratinocyte differentiation or in the mechanism of senescence. Some have even been identified as being involved in the development of skin pathologies such as psoriasis. We will come back to this point in detail later.

Note that the category of non-coding RNAs also includes small interfering RNAs (siRNA — Small Interfering RNA). Like miRNAs, these small interfering RNAs are capable of regulating genes because interference with a specific messenger RNA leads to its degradation and the decrease in its translation into protein.
 

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Exosomes extracted from mesenchymal stem cells: MSC exosomes

Exosomes extracted from mesenchymal stem cells (exosomes MSCs — Mesenchymal stem/stromal cells), present for example in the mesenchyme (tissue) of the embryo, umbilical cord blood, bone marrow or adipose tissue, seem to be the healthiest for therapeutic use. These MSCs exosomes can also be sterilized while MSC stem cells cannot. MSCs exosomes are considered safe in the field of stem cell treatments. MSCs exosomes have been used as alternatives to MSCs stem cell treatments in numerous areas: neurological, cardiovascular, immune system, renal, musculoskeletal, hepatic, respiratory, vision, skin diseases, and cancers.
 

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The origin of MSC exosomes plays a role in the quality of the therapeutic effect

MSC stem cells have the properties of differentiation and self-renewal through cell division. These stem cells can be extracted from several types of tissue or fluids: adipose tissue, bone marrow, dental pulp, placenta, umbilical cord, synovial fluid, amniotic fluid or even Wharton jelly.

Depending on their origin, MSC stem cells can be differentiated into various types of cells such as adipocytes (present in adipose tissue), chondrocytes (found in cartilage), osteoblasts (young bone cell) and myocytes (muscle fibers). MSCs also have immunomodulatory properties that regulate various cells that play a role in the immune response. That's why MScs have been studied so fervently over the past decade.

Since the characteristics and functionalities of MSCs vary according to their origin, it follows that the same is true for the exosomes that are extracted from them. Research has shown, for example, that exosomes derived from adipose tissue stem cells (MSC-ASC type exosomes) may have a therapeutic effect on Alzheimer's disease but would have contradictory benefits on cancers. The origin of the exosomes therefore plays a role in the type of disease targeted.
 

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The hair cycle

The hair life cycle is a dynamic and complex process, composed of a phase of rapid growth that lasts for a long period of time (anagen phase, lasting 2 to 8 years), a phase during which follicles regress (catagen phase, lasting 2 to 3 weeks) and a resting phase (telogen phase, lasting approximately 3 months). When a new anagen phase is re-triggered, a new follicle is created and expels the old follicle (we also speak of an exogenous phase) :

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Arthurandsam

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Hair anatomy

A mature follicle (anagen) can be divided into a visible upper portion and a lower portion that is perpetually remodeled during cycles. It is in the lower part that the factory that controls the formation of the hair is located, in the bulb. This is where the keratinocyte cell matrix (composed of amplified cells derived from follicle epithelial stem cells) and the hair pigmentation unit are found. When this matrix is activated, cells proliferate and differentiate in the region that surrounds it. Their number then determines the size of the bulb and the diameter of the hair.

When matrix cells stop proliferating and differentiating, they will have generated various cell lines in the hair shaft and inner root sheath (IRS). The outer epithelial sheath (ORS — Outer Root Sheath) is an extension of the epidermis and contains a large number of cells that play specific roles in the life of the follicle.

The bulb also contains the dermal papilla, which consists of mesenchymal cells called dermal papilla fibroblasts. Dermal papilla fibroblasts generate signals in the matrix causing the proliferation, migration, and differentiation of cells in this matrix, which are essential for hair follicle growth. The dermal papilla regulates the size of the bulb, the diameter of the hair and its length as well as the duration of the anagen phase.


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Embryo follicle development

The essential prerequisite for the development of a follicle is molecular communication between the cells of the epidermis and those of the underlying mesenchymal tissue of the dermis. These communications are organized according to transduction channels, and there are several types. Signal transduction is the mechanism by which a cell responds to the information it receives, through chemical agents or other signals (voltage, etc.). It controls a cascade of secondary signals, internal to the cell (“signalling”) or external (for example an action on other cell types via interleukins), and internal cellular processes (metabolism, cell cycle, mortality, etc.).

The vast majority of transduction pathways involve the binding of molecules (called ligands) to cell receptors that trigger events within the cell. Among the molecules that can bind to receptors, we find for example a large number of growth factors (proteins that regulate the manufacture or growth of certain cells), cytokines (a substance secreted by cells of the immune system so that they can communicate with each other), neuropeptides (hormone that acts as a neuromodulator) and hormones (chemical substance made by a gland and that act on the development of an organ), which in part are produced by the follicle itself. The follicle is therefore very sensitive to these different molecules :

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Among the molecules secreted by the transduction pathways that interest us in the case of communication between epidermal cells and dermal mesenchymal tissue cells, there are molecules involved in the following signaling pathway families :

  • Wnt/Wingless: Wnt signaling pathways are a group of signal transduction pathways that start with proteins that transmit signals into cells via transmembrane receptors. The clinical importance of this pathway has been demonstrated by mutations that lead to a variety of diseases, including breast and prostate cancer, glioblastoma, type 2 diabetes, and others.
  • Beta-catenin is a protein that plays a very important role in this transduction pathway and plays an essential role in alopecia. Indeed, we will see later that people suffering from alopecia have a defect in canonical Wnt/beta-catenin signaling.
  • Hedgehog: This signaling pathway is activated via ligand binding to Patched receptors, a large transmembrane protein. Involved in embryogenesis and morphogenesis, it plays a major role in the orientation of stem cells and their differentiation. Several pathologies are associated with the mutation of the hedgehog gene in animals, including limb abnormalities and polymalformative syndromes.
  • TGF-beta/BMP (Transforming Grow Factor-beta) : The transforming growth factor beta signaling pathway is involved in numerous cellular processes both in the body and in the developing embryo, including cell growth, cell differentiation, cell migration, apoptosis, cell homeostasis, and other cellular functions
  • BMPs (bone morphogenetic proteins) are morphogenetic bone proteins. It is a family of proteins, such as TGF-BETA growth factor.
  • FGF (Fibroblast Grow Factor) : Fibroblast growth factors form a family comprising 23 proteins identified to date in humans (FGF1, FGF2,... FGF23). These are proteins that activate the migration and multiplication of target cells. These factors are generally secreted by fibroblasts. The most important role of fibroblasts, connective tissue cells, is to maintain the extracellular matrix of connective tissue, and to repair injuries due to trauma. The mechanism of action of FGFs on their target cells involves their binding to specific receptors on the cell surface. These FGF receptors (FGFRs) are a group of molecules capable of receiving signals from outside, transmitting them inside the cell, and inducing numerous secondary cellular messages.
  • TNF (Tumour Necrosis Factor): Tumor necrosis factors form a superfamily of proteins whose type member is TNF-alpha. This cytokine is involved in the maintenance of immune system homeostasis, inflammation, and host defense. They are transmembrane proteins.
  • Notch: The Notch signaling pathway will make it possible to determine the lineage of a cell. Notch signaling is a juxtacrine signaling pathway between two cells in contact with each other. The pathway is activated by the binding of the Notch receptor with one of its ligands, the Delta and Serrate proteins, which are themselves transmembrane proteins.
The formation of a hair, which occurs mainly during embryonic life, is induced by a specific combination of these different signals. It includes three phases: induction, organogenesis, and cytodifferentiation.


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The induction phase (steps 0 and 1) :

  • At the beginning (stage 0), before the visible formation of the placode (cell outline), the interaction between the epidermis and the underlying dermis causes the activation of follicle formation by various signaling pathways, including Wnt/-catenin, which surpass follicle inhibitors.
  • During stages 0 and 1, the placode becomes visible and underlying growth is initiated.
  • From that moment on, the dermis condenses and the future dermal papilla forms.
  • During these placode formation stages, morphogenetic bone proteins (BMPs) must be under-regulated by BMP antagonists in order for the placode to grow. These inhibitors include the noggin protein,
During steps 2 to 8 (organogenesis and cytodifferentiation phases), the orientation of the hair is defined and the bulb develops.

During the hair formation phase, we can therefore notice some proteins that are over-represented or, on the contrary, under-represented. For example:

  • The increase in beta-catenin and Noggin/Lef-1 proteins during placode formation:
    • The protein beta-catenin is created via the canonical Wnt signaling pathway;
    • The noggin protein and the LEF1 protein (Lymphoid enchancer-binding factor 1). These proteins belong to the family of TCF/LEF genes involved in the Wnt signaling pathway, in particular during stem cell development. Genes in this family encode transcription factors. They also play a role in cancer and diabetes.
  • This increase is accompanied by the decrease in beta-catenin inhibitors: Dkk1,2,4 and BMP2,4 and 7.
  • The increase in Wnt10a and Wnt10b, LEF1, FGFs, TGF-beta2 proteins during the proliferation phase of hair stem cells in the epidermis;
  • The increase in PDGF-A and Shh proteins (which is one of the three proteins involved in the Hedgehog signaling pathway) during the dermal papilla formation phase;
Although hair development occurs mainly during embryonic life, the signaling pathways that are implemented there also play a very important role in the regeneration of hair in adults during their life cycle. Indeed, these signaling pathways regulate interactions between epithelial cells and mesenchymal cells in follicles. For example, there are the hedgehog pathways (via the ssh protein), notch, TGF-beta and the Wnt pathway.
 
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Arthurandsam

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The follicle cycle

Once the first postnatal follicle has formed, it begins its life cycle:


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(APM: arrector pili muscle; DC, dermal condensate (green); DP: dermal papilla (green); HS: hair shaft (brown); IRS: inner root, sheath (blue); MC: melanocytes; ORS: outer root sheath; SC: sebocytes (yellow); SG: sebaceous gland)

During the anagen phase, the keratinocyte matrix resulting from progenitor cells (of transient amplification) derived from epithelial stem cells from follicles contained in the bulb, proliferate intensively and generate numerous epithelial cells. This proliferation is activated by the signals generated from the cells of the dermal papilla.

During the catagen phase, two-thirds of the base of the follicle regresses mainly through apoptosis (self-destruction) of the keratinocyte cells composing the outer layers of the hair, while the stem cells contained in the bulb escape apoptosis. The epithelial duct also narrows, which separates the dermal papilla from the bulb.

When the catagen phase is over, the follicles go into dormancy (telogen phase). This phase usually lasts longer than 3 weeks and gets longer with age. During this phase, the dermal papilla remains close to the bulb, which allows a very strong interaction between the stem cells contained in the bulb and the dermal papilla. The dermal papilla plays an essential role in activating stem cells and initializing a new cycle.

Although there are strong correlations between the molecules that will participate in the development of a follicle during the embryonic stage and those that will participate in the renewal of a follicle during the hair life cycle, some differences exist. For example, some members of the TGF-beta signaling pathway and certain growth factors have opposite functions during embryonic follicle development and renewal: while they stimulate their embryonic development, it stimulates the catagen phase in mature follicles. Other signals, such as those from the notch or VDR signaling pathways (which are controlled by vitamin D), are not essential for follicle development but are essential to induce the anagen phase.

Some molecular mechanisms that dictate the dynamics of these different phases are known. For example, the anagen phase is activated by Wnt/beta-catenin, BMP antagonists, and Shh. However, proteins like TGF-beta1, TFG-beta2, BMP2/4, and TNF-alpha generate the catagen phase.

IGF-1 growth factors (Insulin-like Growth Factor One), HGF hepatocytes, vascular endothelium VEGF growth factors (Vascular Endothelial Growth Factor) and FGF-type growth factors contribute to the extension of the anagen phase.

Another molecule that controls the transitions between the anagen and catagen phases are vitamin D receptors.

The telogen phase coincides with major changes in gene activities. Indeed, certain regulatory proteins, such as estrogen receptors, are over-represented during this phase. The telogen phase is therefore not a completely resting phase. It may even be a crucial phase in controlling hair cycles.

In fact, the telogen phase can be divided into a phase that is refractory to stimuli that induce follicle growth and characterized by the overregulation and activation of BMP2/4 proteins and a complementary phase where bulb stem cells become very sensitive to factors inducing the anagen phase. During this phase, the BMP signaling pathway is switched off while the Wnt/beta-catenin signaling pathway is activated and reaches an optimal level of activity at the beginning of the anagen phase.

During the telogen phase, it was seen that the dermal papillae are just below the bulb. When a fairly high concentration of stem cell activators is reached, the anagen phase resumes.

Certain molecules that play a role in the balance between whether stem cells are at rest or not have been identified. While stem cells seem to be kept at rest by the activation of the BMP signaling pathway in combination with the inhibition of the Wnt pathway by the TCF3 and DkkS proteins, they are activated by the inhibition of the BMP pathway and the activation of the Wnt pathway with stabilization of the protein beta-catenin :

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During the onset of the anagen phase, the bulb gradually moves away from the dermal papilla and the epithelial stem cells go into dormancy. However, the Wnt/beta-catenin signaling pathway remains active thanks to the progenitor cells (transient amplification) contained in the hair matrix. These cells also stabilize beta-catenin throughout the anagen phase. A little higher in the matrix, these cells stop proliferating and initiate terminal differentiations into different epithelial cell lines.

Numerous molecules that regulate the formation of the internal and external epithelial sheath have been discovered. For example, GATA3, Cutl1, and BMPs participate in the formation of the inner epithelial sheath, Sox9 and SHH participate in the formation of the outer epithelial sheath, and Wnt/-catenin, vitamin D receptors, Notch, BMPs, and Foxn1 participate in hair shaft formation. In contrast, TCF3 is a general inhibitor of all epithelial cell lines.
 
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Arthurandsam

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Experiments have shown that MSC-ASC exosomes cause hair growth

Exosomes contain microRNAs that, when delivered to other cells, will affect their behavior. The microRNAs in exosomes vary depending on the stem cells from which they come. Some microRNAs will increase follicle proliferation while others will tend to slow them down.

The regulation of follicle growth is regulated by a paracrine mechanism in which exosomes from dermal papilla cells play an essential role.

But in addition to this type of cell, ASCs located in the fatty tissue under the dermis can also play a role in the hair cycle because ASCs differentiate into mature adipocytes that surround the hair during the transition between the telogen - anagen phase.

Numerous studies have also shown that ASCs have an effect on hair growth by increasing the proliferation of dermal papilla cells. This is because interactions between ASC cells and dermal papilla cells are controlled by various mediators that activate the telogen—anagen transition. In particular, as has been seen, the Wnt/-catenin signaling pathway and the IFG-1 growth factors produced by dermal papilla cells. Exosomes derived from MSC stem cells carry a large number of growth factors and activators of Wnt/beta-catenin signaling.
 

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The proteins contained in the exosomes of the ASCE+ HRLV product

Below are listed some proteins transported by the exosomes contained in the ASCE+ HRLV product. It can be noted that a large number of them participate in the proper development of follicles:

  • sh-Oligopeptide-1: EGF growth factor, sh-EGF
    • Stimulates cell growth, differentiation and proliferation
    • Participate in the extension of the anagen phase
  • SH-oligopeptide-2: IGF1 growth factor
    • Stimulates cell proliferation and wound healing
    • Participate in the extension of the anagen phase
These two groups of proteins work in synergy to promote the proliferation of keratinocyte (cells that make up the outer layer of the skin).

  • sh-oligopeptide-4: Thymosin-beta-4 peptide
    • Stimulates the growth, migration, and proliferation of follicle stem cells between the bulb and the dermal papilla during the telogen phase, which promotes follicle growth and accelerates the transition to the anagen phase.
  • sh-Polypeptide-3: KGF growth factor
    • It affects the proliferation and differentiation of various cells because this protein is part of the FGF family. This family is involved in the growth of the follicle when it is born.
  • sh-Polypeptide-4: SCF peptide (Stem cell factor)
    • An oxytokin that controls the lifespan, growth and differentiation of hematopoietic stem cells.
    • Facilitates the generation of melanin and hair growth factors
  • sh-Polypeptide-9: VEGF growth factor
    • Improves cytokine division and proliferation
    • This protein plays a crucial role in vascularization around follicles during hair cycles.
    • Improves the effect of FGFs
  • sh-Polypeptide-13: Noggin
    • Stimulates hair growth and inhibits hair depigmentation.
    • Participates in placoid formation.
    • BMP4 antagonist (which inhibits follicle development)
    • Improves cell proliferation
  • sh-Polypeptide-8: Platelet-Derived Growth Factor (PDGF)
    • Intervenes in skin growth and wound repair
    • Stimulates the production of collagen and elastin and improves skin elasticity
  • sh-Polypeptide-1: bFGF growth factor (Fibroblast Growth Factor)
    • Regulates skin renewal
    • Stimulates the multiplication and division of extracellular matrices (dermal fibroblasts), keratinocytes, and many other types of cells
    • Stimulates hair growth by increasing collagen and elastin synthesis and restricts hair discoloration
    • Controls the spread, migration, and differentiation of epidermal cells by improving blood circulation and nutrient delivery
    • In interaction with EGF and IGF-1, boosts hyaluronic acid production and improves skin quality
  • sh-Polypeptide-11: aFGF growth factor
    • Stimulates the proliferation of fibroblast and keratinocyte cells
    • Improves skin regeneration
    • Improves blood circulation and vascularization by stimulating endothelial cell proliferation.
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Alopecia and DHT

We have seen that hair growth is governed by various endogenous factors, such as intracellular and inter-cellular signaling. For example, BMP and TGF signaling (especially TFG-beta1 and TGF-beta2 factors) are known to inhibit the induction of the anagen phase, while the Wnt10b protein stimulates the transition from the telogen phase to the anagen phase. In the same way, FGF-type growth factors induce the anagen phase. Finally, other signaling pathways, when they are overactivated or on the contrary underactivated, play a role in the adult hair cycle :

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One of the main consequences of alopecia is the gradual reduction in the duration of the anagen phase. The follicles pass prematurely from the anagen phase to the catagen phase. Another reason is the miniaturization of hair, a process during which terminal solid hair is transformed into fluffy hair.

The influence of the androgen hormone DHT (dihydrotestosterone) on hair loss is well known. Androgens, including DHT, enter follicles through the capillary network in the dermal papilla. They then bind themselves to the androgen receptors contained in the cells of the dermal papilla, and activate or on the contrary deactivate certain genes. However, in people with alopecia, the dermal papilla cells contained in the follicles secrete the proteins TGF-beta1 and TGF-beta2. These inhibit the growth of epithelial cells in response to androgens.

In the body, testosterone is converted to DHT by the 5-reductase enzyme. However, experiments have shown that dermal papilla cells derived from follicles in the front of the scalp (which contain fewer ARA70beta/ELE1beta transcriptional androgen coactivating receptors but more type II 5-alpha-reductase androgen receptors than cells from occipital dermal papillae - which “do not fall”), are known to secrete inhibitory autocrine factors that affect cell growth. This shows that DHT alters autocrine and paracrine processes and plays an essential role in hair loss, more than other androgens.

Several studies have shown that under the effect of DHT, the DKK1 gene (dickkopf 1) is over-regulated in the cells of the dermal papillae of people with alopecia. This over-representation also has an impact on the growth of keratinocytes in the outer epithelial sheath.

We will see in the next section that the Dkk1 protein plays a very important role in inhibiting the proper functioning of the canonical Wnt/beta-catenin transduction pathway. The role that DHT plays on our hair can therefore be summarized by the following short diagram:

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The importance of the Wnt/beta-catenin signaling pathway on hair growth

As we have seen: there are numerous endogenous factors that influence hair growth, including molecules involved in intercellular and extracellular signaling pathways. Activation of the anagen phase is inhibited by the activation of BMP signaling proteins and TGF-beta growth factors. In contrast, the Wnt/beta-catenin and hedgehog (SHH) signaling pathways stimulate hair growth and entry into the anagen phase.

We will see that in the case of the Wnt signaling pathway causing hair growth, the protein beta-catenin is stabilized in the cytoplasm of the cell and translocated into the nucleus, where it acts with proteins of the TCF/LEF family as a transcription factor. In this case, we say that the Wnt path is canonical. This results in the activation of genes that promote the activation of the anagen phase and therefore the growth of a new hair.

The TCF/LEF (T-cell factor/ Lymphoid Endancer Factor) family is a group of genes that encode transcription factors. They are involved in the Wnt signaling pathway, especially during the embryonic phase and stem cell development, but they also play a role in cancer and diabetes. TCF/LEF factors recruit beta-catenin as a coactivator in order to increase the probability of transcription of a specific gene. On the contrary, they can also recruit a member of the Groucho family as corepressors (and therefore decrease the probability of gene transcription). When the protein -catenin is not translocated into the nucleus but destroyed in the cell's cytoplasm, it is the Groucho family protein that is recruited by TCF/LEF factors. In this case, we say that the signaling pathway is not canonical. Unfortunately, this is what happens to people with alopecia.

Numerous studies have shown that multiple proteins from the Wnt family are involved in the proper functioning of the hair cycle and its regeneration. For example:

  • Injections into the dermis of products containing Wnt1a proteins derived from MSC stem cells extracted from the marrow (B-MSC, Bone Marrow Mesenchymal Stem Cells) on mice accelerate the transition from the telogen phase to the anagen phase, increase the quantity of hair, and increase the expression of genes related to hair formation, such as LEF1.
  • The Wnt10b protein improves the differentiation of epithelial cells to form the hair shaft and the internal epithelial sheath and the transition from the telogen phase to the anagen phase.
  • Injections of extracellular vesicles from macrophages (cells belonging to white blood cells) that contain large quantities of Wnt3a and Wnt7b significantly increase the proliferation of dermal papilla cells as well as the diameter of the hair shaft.
  • On the other hand, the Wnt5a protein, one of the proteins expressed in the non-canonical Wnt pathway, thwarts the functions of the canonical Wnt pathway. Injections of adenoviruses over-expressing Wnt5a into mice resulted in an extension of the duration of the telogen phase and an attenuation of the entry into the anagen phase, all associated with a decrease in the expression of -catenin. The decrease in cell proliferation induced by Wnt5a could then be reversed by the addition of Wnt3a.
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Among all the associated signaling pathways in the hair cycle, many studies assume that that of Wnt/beta-catenin is the most important because DHT, which is over-regulated in alopecia, alters this signaling pathway. The main inhibitors of this signaling pathway are the proteins DKK1, SFRP2 (Secreted Frizzled-Related Protein 2) from the SFRP family and sclerotin (which binds to BMP7).

DKK1 is a powerful inhibitor of the Wnt/beta-catenin transduction pathway and experiments have shown that injections of DKK1 into the mouse scalp activate the anagen — catagen transition. This protein is over-represented in patients with alopecia and DHT interferes with the normal hair cycle in a negative way by activating the catagen phase through the involvement of DKK1.

It is therefore reasonable to postulate that the inhibition of DKK1 is a key to promoting hair growth in people with alopecia. In addition, there are a number of microRNAs that target DKK1 and are involved in signaling pathways related to hair growth.

Let us now see in more detail how the canonical Wnt/beta-catenin signaling pathway works and how it is deteriorated in alopecia.
 
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The canonical Wnt signaling pathway and its deterioration in alopecia

As already mentioned, the canonical Wnt signaling pathway (or Wnt/beta-catenin signaling pathway) is the Wnt signaling pathway that causes accumulation of the -catenin protein in the cytoplasm of the cell and its possible translocation into the nucleus to act as a transcriptional coactivator of transcription factors belonging to the TCF/LEF family.

Let's see in more detail how the DKK1 protein negatively influences this canonical signaling pathway.

At rest, there is beta-catenin in the cell's cytoplasm, which is phosphorylated so as not to accumulate (the phosphorylation process is a chemical modification that consists in adding a phosphate group to a molecule). The destruction complex responsible for this phosphorylation process is composed of the proteins APC (Adenomatous Polyposis Coli), GSK-3beta (Glycogen Synthase Kinase 3beta), CK1alpha (Casein Kinase 1alpha), and PP2A (Protein Phosphate 2A). This destruction complex marks beta-catenin for degradation by the proteasome (proteasomes are enzymatic complexes that are found in the nucleus of cells and whose role is to degrade misfolded proteins — that is, a malformed protein).

During the canonical phase of the Wnt/beta-catenin signaling pathway, the Wnt ligands bind to the FZD receptor (Frizzled) and to the LRP5/P6 co-receptor. This co-receptor is transmembranous: one part is found outside the cell and another in the cell's cytoplasm. As soon as Wnt binds to Frizzled and LRP5/P6, this causes the translocation of the negative Wnt regulator AXN (Axin), which binds to the tail of the LRP5/P6 protein found in the cytoplasm. The PP2A phosphate group of the destruction complex then induces phosphorylation of the tail of the LRP5/P6 protein. This process is followed by the recruitment by phosphorylation of the phosphoprotein DVL (dishevelled), which binds to the frizzled receptor in the endodomain of the cell, which inhibits the activity of GSK-3beta and therefore deactivates the destruction complex. This prevents the protein -catenin from being degraded and sent to the proteasome to be digested. The beta-catenin can then accumulate and translocate in the nucleus, and then bind to the TCF/LEF transcription factors. A transcriptional complex is then formed, and the transcription of a certain number of Wnt genes is then activated.

On the other hand, when the Wnt ligands do not bind to the FZD receptor, beta-catenin becomes unstable in the cytoplasm because it is phosphorylated by GSK-3beta and CK1alpha since the negative regulator AXN is not translocated at the LRP5/P6 tail, and therefore DVL is not recruited. GSK-3beta is then not inhibited and the transcriptional complex is therefore not formed in the nucleus. In this case, the transcription factor TCF/LEF recruits a member of the Groucho family as a corepressor.

Studies have shown that when DKK1 proteins are over-regulated, they bind to LRP5/P6, which suppresses the Wnt signaling cascade, and inhibits follicle development, and therefore hair growth. The DKK1 protein is therefore a key factor in the development of alopecia.

Some studies also seem to show that the SFRP2 protein could inhibit the Wnt/beta-catenin signaling pathway, but this has not yet been proven. Indeed, treatment with human SFRP2 recombinants on dermal papilla cells extracted from patients undergoing transplantation considerably increased canonical Wnt signaling.


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Arthurandsam

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DHT level in correlated with DKK1 level

Studies have shown that the level of DHT and DKK1 are correlated. In the body, 5 alpha reductase (5alphaR) converts testosterone to DHT. This androgen hormone plays an inhibitory role on the growth of outer epithelial sheath cells, which induces a disturbance on normal hair growth. Experiments have also shown that anti-DKK1 drugs greatly reduce the inhibition of the growth of outer epithelial sheath cells. In addition, the concentration of DKK1 is higher in the scalps of people with alopecia. Finally, studies have shown that DKK1 disrupted by DHT stimulates the apoptosis of keratinocyte cells.

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Some molecules inhibit the DKK1 protein

Some studies have looked at the effect of certain molecules on the canonical Wnt pathway. A study observed the impact of morroniside, a natural molecule found in male dogwood (sometimes called cornflower or fuselaria), on the canonical Wnt signaling pathway in cells of the outer epithelial sheath. The results show an increase in cell proliferation as well as an increase in β-catenin. Another concurrent study showed that this treatment acted on DKK1.

Another natural compound, vitexin (found for example in Passionflower), significantly increases the proliferation of dermal papillae in follicles. Again, vitexin has been shown to over-regulate β-catenin and substantially decrease the level of DKK1.

Ginseg extract also caused keratinocyte proliferation in the outer epithelial sheath, inhibited apoptosis, and revealed the effect of DKK1.

Another natural compound that activates the WNT/beta-catenin and SHH pathway while inhibiting the TGF-beta/SMAD and BMP pathways in follicles, and consequently induces hair growth, is costunolide. It is found in the Indian costume.

The essential components of Ginkgo Bilboa, Ginkkolide B and Bilobalide, are also known as molecules that can help with hair growth. These molecules are associated with the activation of the Wnt/beta-catenin pathway via the inhibition of DKK1 gene expression.

The antidepressant tianeptine is known to have an impact on hair diameter, and therefore hair growth, by inhibiting DKK1. It also slows the transition from the anagen phase to the catagen phase in people with alopecia.

Finally, minoxidil also under-regulates the production of DKK1 and TGF-beta in keratinocyte cells.

It can therefore be seen that many studies show the influence of the inhibition of DKK1 in hair growth.
 

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The role of microRNAs in modulating hair growth

A number of microRNAs are involved in hair growth and the deficiency of the canonical Wnt pathway impacts this growth. The idea is therefore to use certain microRNAs to regulate this pathway. I have previously focused in this report on studying the Wnt pathway, which is the most important pathway in the field of hair growth, but as we have already mentioned before, there are many other signaling pathways involved in the hair growth process.

In what follows, I focused in particular on the microRNAs that are found in the exosomes of MSC stem cells and derived from dermal papillae.

A study has shown that exosomes derived from dermal papilla cells contain nearly 200 microRNAs. The 34 micro-RNAs that are over-represented or on the contrary under-represented are the following: let-7b-5p, let-7c-5p, let-7f-5p, let-7g-5p, miR-145-3p, miR-122, miR-126-3p, miR-126-5p, miR-128-3p, miR-133a-3p, miR-145-5p, miR-195-3p, miR-1919p 3b-5p, miR-199c-3p, miR-199c-5p, miR-200a, miR-200c, miR-214-3p, miR-222-3p, miR-22-5p, miR-24-3p, miR-27b-3p, miR-320-3p, miR-330-5p, miR-34a, miR-423-5p, miR-4a-49a 5p, miR-451-5p, miR-499-5p, and miR-99a-5p.

This study showed that these microRNAs are involved in 40 biological functions, in particular in those related to intercellular communication intervening in follicles, such as:

  • The Wnt signaling pathway
  • Axonal guidance: a branch of neurodevelopment,
  • Glycosylphosphatidylinositol: GPI — which is a glycolipid that allows the anchoring of various molecules, in particular proteins to cell membranes, and of the red blood cell in particular,
  • Endocytosis: mechanism for transporting molecules, or even particles, to the inside of the cell,
  • Fatty acid metabolism: consists of various metabolic processes involving or closely related to fatty acids, a family of molecules classified in the category of lipid macronutrients,
  • RAS-MAPK signaling pathway: The RAS/MAPK pathway is an intracellular signaling pathway that plays an important role in the regulation of cell proliferation, survival, differentiation and migration, as well as angiogenesis (the process of growth of new blood vessels from pre-existing vessels).
  • The JAK/STAT signaling pathway: a chain of interactions between proteins in a cell and is involved in processes such as immunity, cell division, cell death, and tumor formation.
These 34 microRNAs potentially target 33,055 genes. The expression of some of these genes will have the consequence of increasing or decreasing the number of proteins coded. Thus, some microRNAs have a positive effect on hair growth while others will have a negative effect.

This same study then studied the interaction of these 34 microRNAs with 29 genes involved in hair growth in order to identify the most represented microRNAs:

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The 9 representatives are the microRNAs miR-200b, miR-200c, let-7b-5p, miR-122, miR-200a, let-7g-5p, let-7c-5p, miR-22-5p, and let-7f-5p. Among them, the main microRNAs identified as targeting genes related to follicle growth are miR-22-5p and let-7b-5p. The study then showed experimentally that miR-22-5p inhibits the proliferation of follicle stem cells by targeting the LEF1 gene, which is involved in the canonical Wnt pathway.

Of course, other microRNAs possibly derived from other exosomes originating from different stem cells may have an impact on hair development. It is known that exosomes derived from MSC stem cells have a beneficial effect on hair growth. It contains the following microRNAs:

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Among the microRNAs that have an impact on the regulation of DKK1, we find for example:

  • miR-1-3p: When its level decreases, DKK1 increases
  • miR-34-a: As its level increases, DKK1 decreases
  • miR-103-3p: As its level increases, DKK1 decreases
  • miR-146A: When its level increases, DKK1 decreases (found in MSC exosomes)
Among the microRNAs known to have an impact on hair growth, we find for example:

  • miR-let-7a: Action to regulate hair growth by targeting c-Myc, IGF-1R and FGF5.
  • miR-let-7b: Action to regulate hair growth by targeting EDA.
  • miR-21: Action to regulate hair growth by targeting CNKSR2, KLF3 and TNPO1.
  • miR-22: Negative action on hair growth by targeting STK40 and inhibiting MEF2-ALP which impairs the proliferation and differentiation of follicle cells (found in MSC exosomes).
  • miR-29a/B1: Negative action because its overexpression induces hair loss by targeting Wnt and BMP.
  • miR-103/107: Positive action by activating PKC signaling which maintains the quality of differentiated cells (quality that decreases with age).
  • miR-130a-3p: Positive action by regulation of TGF-beta; miR-130a inhibits the activation of TFG-beta by targeting the TGFBR1 receptor (found in MSCs).
  • miR-140-5p: Positive action by inhibiting the BMP pathway by targeting BMP7.
  • miR-200: Positive action by regulating the Hippo/Yap pathway.
  • miR-214: Negative action by inhibiting the Wnt pathway.
  • miR-125b and miR-221: Negative action because this microRNA is over-regulated in the dermal papillae of people with alopecia (found in MSCs).
It is interesting to note that even the presence of microRNAs that play a negative role on hair growth independently (such as miR-22 in MSC exosomes) does not prevent the exosomes of these stem cells from acting positively overall. I think that this comes from the fact that their number is lower in proportion to other microRNAs that have a positive effect and that the dynamics on which all microRNAs act is very complex. Compensation phenomena must certainly occur.
 
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What microRNAs are contained in ASCE+ HRLV exosomes?

The question is therefore to know whether the microRNAs contained in the ASCE+ HRLV product can play a beneficial role in the proliferation of follicles, their maintenance in the anagen phase, or even the ability to force the hair to move rapidly from the telogen phase to the anagen phase.

Unfortunately, the exosomes contained in the ASCE+ HRLV product are not exosomes extracted from dermal papilla stem cells, or mesenchymal stem cells. The reason is that it is forbidden to sell such exosomes to the general public in some countries (such as in Europe, the United Kingdom, the US,...), while it can be done in other countries (Korea, Japan,...).

The exosomes contained in the ASCE+ HRLV product come from rose stem cells (the English term is “Rose stem cell exosomes”). In particular, the company ExoCoBio has filed the “ExoScrt” patent allowing this extraction. This makes it possible to make the use of derivatives of MSC exosomes accessible in certain countries where exosomes derived from human stem cells are prohibited. These exosomes have collagen-synthesizing properties and anti-inflammatory properties.
The study of these exosomes has shown that they share 904 microRNAs with those found in humans. Among these micro-RNAs, 5 are in direct relationship with cell proliferation and three (miR-8485, miR-574-5p and mir-1246) play a role in the regulation of Wnt signaling:


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Among the microRNAs identified as having an impact on hair growth, there are a number of interesting ones:

  • miR-1-3p: When its level decreases, DKK1 increases
  • miR-let-7a: Action to regulate hair growth by targeting c-Myc, IGF-1R and FGF5.
  • miR-let-7b: Action to regulate hair growth by targeting EDA.
  • miR-21: Action to regulate hair growth by targeting CNKSR2, KLF3 and TNPO1.
  • miR-29a/B1: Negative action because its overexpression induces hair loss by targeting Wnt and BMP.
  • miR-103/107: Positive action by activating PKC signaling which maintains the quality of differentiated cells (quality that decreases with age).
  • miR-130a-3p: Positive action by regulation of TGF-beta; miR-130a inhibits the activation of TFG-beta by targeting the TGFBR1 receptor (found in MSCs).
  • miR-214: Negative action by inhibiting the Wnt pathway.
Those that are also found in MSC exosomes or dermal papilla stem cell extracts include: miR-let-7a-5p, miR-Let-7b-5p, miR-Let-7c-5p, miR-Let-7f-5p, miR-Let-7g-5p, miR-Let-7i-5p, miR-Let-7i-5p, miR-122-5p, miR-1246, miR-130a-3p, miR-21-5p, miR-214-3p, miR-23a-3p, miR-23b-3p, miR-29b-3p, miR-4488.

Many of the microRNAs carried in rose exosomes also have anti-aging, anti-inflammatory, regenerative and wound-healing properties. I did not study these microRNAs because I focused on those that have an impact on hair, but here is the summary given by the ExocoBio laboratory on ASCE+ HRLV exosomes:


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If you want information on specific microRNAs, you can use the search engine https://www.genecards.org/genecarna. You will be able to have a list of the scientific studies carried out on this microRNA in order to understand its various implications in the human body. For example, a search for miR-130a microRNA gives 288 results:

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Conclusion

The ASCE+ HRLV product therefore seems interesting because it contains a lot of growth factors and microRNAs that have positive actions on hair growth.

However, a few questions remain:

  • The ASCE+ HRLV product contains non-human exosomes... Does this have an impact on the quality of the microRNAs that will be found there?
  • The reassuring point is that many of the microRNAs contained in the product are found in exosomes derived from MSC stem cells or dermal papillae.
  • The less reassuring point is that some microRNAs contained in the ASCE+ HRLV product seem to have a negative impact on hair growth. That said, these microRNAs are also found in the exosomes of MSC stem cells or dermal papillae and numerous studies have proven their effectiveness on hair growth.
I could not find the complete list of microRNAs contained in rose exosomes but the data should normally be public soon. It would be interesting to know whether other negatively or positively acting microRNAs are contained in these exosomes.
 

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Update at +3,5 months : No full head of hair but still on it :rolleyes:

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I would like to take this opportunity to share with you two very interesting new videos dated from May 2023 on exosomes I recently discover if this subject interests you:
  • The first one (with a delicious French accent
    1710625630019.gif
    ) talk about Extracellular vesicles (Exosomes) characterization and therapeutic potential ; very very interesting ! for example, they talk about the difference between stem cell and exosome treatments and the fact that the latter method is much safer... Talk also about Platelets...
  • The second one is some Q&A about exosomes, and they speak about exosomes for hairloss... and you will see that there is no propaganda on this subject, they are rather reserved about this type of treatment for the moment...
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Note: Note: I specify that I do not know at all if exosomes work to fight against hair loss and have any interest... Personally I think that in certain cases it can be beneficial, in particular when the folicles are not dead , but I'm just reporting my experience !

<========================================================================================================>

My point of view : I think that the exosomes currently available to the general public are not yet necessarily "loaded" with specific genetic material (mi-RNA) to treat alopecia... they in fact contain a mixture of mi-RNAs including one some are beneficial and others less (see my web-site: https://suiviexo.000webhostapp.com/index-en.html ... even in exosome derivated from rose stem cell there are interessting mi-RNA wich are in stem cells from humans !)

In particular, exosomes derived from human stem cells are banned in my country and many other countries. This is the reason why we can only use exosomes from, for example, plants... but which still contain interesting genetic material for hair!

The future is in exosome engineering: Researchers are able to "load" into exosomes specific genetic material. All that remains is to find interesting mi-RNAs to fight against alopecia (we already know a large number of them!) and make them accessible to the general public!

And of course, for the moment no scientific study has been carried out by FDA to validate the therapeutic benefits of exosomes as drugs... It will therefore still take many years to "eventually" see them arrive on the market ! At the moment, they are only available as cosmetics!
 
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