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New Strategies for Tissue Regeneration
Have you ever thought that you want to be more like a newt? You might not have thought about it in those terms, but these tiny amphibians have a physical capability that human beings have dreamed about for eons: the capability of regenerating tissue. If we could tap into this capability, the possibilities for medical treatment are limitless. We could regrow an arm, a leg, a hand, repair a heart after a heart attack, or even regrow hair. Two new avenues of scientific research, discussed in an article in the New York Times, might just help us enable human beings regenerate tissue.
The Stanford Approach
For ages, it has been well known that humans do not possess the regenerative powers of lower vertebrates, such as newts and fish, but the reason has been a mystery. The researchers at Stanford University in California, working with mouse muscle cells, have begun to understand the mechanism behind the capability for certain animals to regenerate tissue.
It seems that lower vertebrates have a genetic makeup that allows their cells to multiply when tissue regeneration is needed. Since unchecked cell multiplication can also lead to tumor (cancer) formation, they also have a tumor suppressor gene known as Rb. This gene is naturally inactivated in newts and fish when they start regenerating tissue.
Mammals possess both the Rb gene and a backup, called the Arf gene, which will close down a cancer-prone cell if Rb fails to do so. [...]
The Stanford team shut off both Rb and Arf with a chemical called silencing-RNA and found that the mouse muscle cells started dividing. When injected into a mouse’s leg, the cells fused into the existing muscle fibers, just as they are meant to.
It would appear then, that mammals, including humans, have regenerative capabilities normally programmed into their DNA, but over hundreds of millions of years these capabilities have been suppressed so that the more important function -– that of cancer prevention -– could operate. To clone human tissue, one would theoretically just need to deactivate the suppressor genes, but in a way that would not put the person at an increased risk of developing cancer. Of course, these genes have not yet been identified in man, nor is it known if they even exist.
The UCSF Approach
A second, but very different, approach to tissue regeneration has been taken up by Dr. Deepak Srivastava and his team at the University of California, San Francisco. Based on work by Japanese scientist Shinya Yamanaka, Dr. Srivastava successfully converted ordinary tissue cells (fibroblasts) of the mouse heart into heart muscle cells:
[Dr. Yamanaka] showed three years ago that skin cells could be converted to embryonic stem cells simply by adding four proteins known to regulate genes. Inspired by Dr. Yamanaka’s method, Dr. Srivastava and his colleagues selected 14 such proteins and eventually found that with only three of them they could convert heart fibroblast cells into heart muscle cells.
To make clinical use of the discovery, Dr. Srivastava said he would need first to duplicate the process with human cells, and then develop three drugs that could substitute for the three proteins used in the conversion process.
The drugs could then be injected into damaged areas of the heart to repair the cardiac muscle cells following a heart attack. By using heart fibroblasts to produce cardiac muscle cells, rather than using embryonic stem cells, it is possible that risk of unwanted tumor formation, often noted with stem cell therapies, can be avoided.
It is not a stretch to assume that if scientists can undo the inability of animals to grow heart muscle or limbs, we might someday be able to genetically reverse the inability of a bald person to grow hair.
Hair Cloning Shows Promise in New Stem Cell Study
Alopecia areata is an auto-immune disease that causes hair loss that ranges from small circular areas on the scalp to extensive or even total baldness. When extensive, it can be a socially debilitating disease, and it can be particularly difficult when those suffering are children.
When alopecia areata is localized, i.e. there are a limited number of bald patches, the condition often responds well to cortisone injected directly direct into the scalp. When the condition is more extensive, current treatments do not have a high rate of success. A new study, using hair cloning therapy to regrow hair, shows promise for all individuals suffering from the disease.
The study — conducted by Marwa Fawzi, a dermatologist at the University of Cairo Faculty of Medicine, and reported on Bloomberg.com — used stem cells from the scalps of eight children with alopecia areata to regenerate their own hair:
The Cairo researcher took small amounts of skin from the scalps of the children, isolated the hair follicle stem cells that stimulate hair production, and grew them in the lab, increasing the number of cells. After one month, she put the cells back into the scalps of the children, with numerous injections across the bald areas of their heads.
Laminin-511 Stimulates Dermal Papilla for New Hair Growth
Jing Gao, Mindy C. DeRouen, Chih-Hsin Chen, Michael Nguyen, et. al.
Genes & Development 22:2111-2124, 2008
The growth of a hair follicle from its developmental cell stage to a hair bearing follicle is through an interactive process between epidermal cells and those of the dermal papilla. It was found that Laminin-511 is instrumental in facilitating this process.
It has been felt that the extra-cellular protein Laminin is critical to both adhesion and the signaling process in hair development; however, the mechanism is not fully understood.
Through this study, it was shown that the signaling pathways introduced by the administration of noggin and sonic hedgehog alone were insufficient to develop a hair follicle. When Laminin-511 protein was introduced to the tissue culture, the dermal papilla developed. When the protein was inhibited, hair follicle growth again ceased. This information supports prior studies suggesting that Laminin is critical in the early stages of follicle cell development and is required for continued follicle development and growth.
This study reaffirms in vitro and in vivo studies in mice, the importance of Laminin-511 in the formation of dermal papilla to promote the development of more organized dermal papilla cells and the hair follicle development. It also suggests that there is a reciprocal mechanism between the signaling pathways of noggin and sonic hedgehog with Laminin-511.
Strategies for Follicular Cell Implantation
by Jeff Teumer, PhD
Hair Transplant International Forum, Volume 18, Number 3, May/June 2008
Follicular cell implantation (FCI) is based on the ability of the dermal papilla (DP) cells, found at the bottom of hair follicles, to stimulate new hairs to form. DP cells can be grown and multiplied in culture, so that a very small number of cells can produce enough follicles to cover an entire bald scalp.
In order to produce new follicles, two types of cells must be present. The first are Keratinocytes, the major cell type in the hair follicle, and the second are dermal papillae cells (DP) which lie in the upper part of the dermis, just below the hair follicle. It appears that the DP cells can induce the overlying keratinocytes to form hair follicles. There are a number of proposed techniques for hair regeneration that use combinations of cells that are implanted in the skin. The two major techniques involve either transplanting dermal papillae cells by themselves into the skin, or implanting them with keratinocytes. These techniques can be used with or without an associated matrix used to help orient the newly forming follicles.
Biologists Make Skin Cells Work Like Stem Cells
By Nicholas Wade
New York Times, 6/7/2007
Summary:
A major advance in regenerative medicine has recently been announced. A new technique, which can convert adult skin cells into embryonic form, has been successfully performed on interbred mice by Dr. Shinya Yamanaka of Kyoto University. The technique, if adaptable to human cells could allow new heart, liver, or kidney cells to be regenerated from simple skin cells. This tissue could potentially replace organ tissue that has been damaged due to disease. As this tissue would be formed from the patient’s own skin cells, it would not be subject to rejection by the patient’s immune system.
Hair Follicle Regeneration
This study demonstrates that after wounding the skin of an adult mouse, an embryonic-like change in the epidermal cells outside of the hair follicle stem cells can be induced to form new hair follicle stem cells. In other words, these cells originate from epidermal skin cells in the wound, but then are able take on the characteristics of hair follicle stem cells and actually produce hair. These regenerated hair follicles establish a stem cell population that can produce a hair shaft and continue through all stages of the follicular cycle. The research suggests that these regenerated hair follicles grow new hair through the introduction of Wnt proteins.
The technology, developed at the University of Pennsylvania School of Medicine, has been licensed by Follica Inc. a privately held medical device company.
Reference: Hair Follicle Regeneration in Adult Mouse Skin After Wounding
Ito, M., et al. Nature 447, 316-320, May 17, 2007
U.K. Invests in Hair Cloning Research
The British Government has awarded Intercytex a grant to automate the production of their new hair regeneration therapy. Intercytex is a cell therapy company that develops products to restore and regenerate skin and hair. Intercytex has partnered with a private company, The Automation Partnership (TAP), to develop an automated manufacturing process for their novel hair multiplication treatment.
Hedgehog and Hair Growth
Curis, Inc., a drug development company, has published data showing the effectiveness of a proprietary Hedgehog pathway activator to stimulate hair growth in adult mice. The study shows that a topically applied small molecule agonist of the Hedgehog signaling pathway can stimulate hair follicles to pass from the resting stage to the growth stage of the hair cycle. The Hedgehog agonist produces no other noticeable short or long-term changes in the skin of the mice.
This study also demonstrated that the Hedgehog agonist is active in human scalp in vitro as measured by Hedgehog pathway gene expression. The results suggest that topical application of a Hedgehog agonist could be effective in treating hair loss conditions, including male and female pattern genetic hair loss.
Preliminary results were presented at the American Academy of Dermatology (AAD) in February 2005. This work was based on a study In 2001 by Sato et. Al. who showed that the Sonic hedgehog gene is involved in the initiation of hair growth in mice1.
Reference: 1. Sato N., Leopold PL, Crystal, RG. Effect of Adenovirus-Mediated Expression of Sonic Hedgehog Gene on Hair Regrowth in Mice With Chemotherapy-Induced Alopecia. Journal of the National Cancer Institute, 2001, Vol. 93, No. 24.
Hair Cloning: Hope for Bald Baby Boomers
An English based company called Intercytex has claimed some success in its research on hair cloning with its first testing in humans. This technique is similar to the one initially proposed by Dr. Colin Jahoda and published in 1999. Download the Article 
The idea is that certain cells (called fibroblasts) found at the bottom of hair follicles can be separated from the follicles after they have been removed from the scalp, and then be used to form new follicles.
The way this works is as follows: A few hair follicles at the permanent area from the back of the scalp (the area that does not bald) are removed. In a lab, the germinative cells at the base of the follicle are dissected off and placed in a Petri-dish. They are then incubated in a special medium and allowed to multiply thousands of times.
These cultured cells are then injected into the balding area of the scalp where they induce complete hair follicles to form. In contrast to traditional hair transplants, where the doctor is limited by the patients finite donor supply and hair is literally just moved around (from the back to the front), in hair cloning, there will be an actual increase in the total number of hairs on a person’s head.
Initial testing involved 7 male volunteers that were suffering from androgenetic alopecia (common baldness). After the process, 5 of them showed an increased amount of hair. Fortunately, there were no complications, such as skin inflammation or tissue rejection. However, the test area was small and volunteers only grew a little hair.
Towards the middle of next year, additional patients will be tested using a greater number of cloned cells, so that a larger area of the scalp could be covered. The researchers speculate that this new cloning technology may be on the market in as soon as 5 years.
The researchers speculate that in the distant future, traditional hair transplants may not be needed at all. Instead, as patients start to thin, they could come to the clinic on a regular basis for injections of their own cells to stimulate the growth of new follicles and stop the impending balding – a sort of hair maintenance.
Reference: The Plain Dealer, Tuesday, November 15, 2005. “Hope grows for bald baby boomers,” Malcolm Ritter, Associated Press.
Skin Cells Substitute for Embryonic Stem Cells in Cloning Research
The advantage of using embryonic stem cells in cloning research, organ transplantation, and in finding cures for disease, is that these cells are basically “unprogrammed.” This means that the stem cell has not yet determined what it will grow to become so, in theory at least, scientists can manipulate them into becoming anything that they are programmed to be.
Two teams of scientists working independently (Kazutoshi Takahashi and Shinya Yamanaka at Kyoto University, Japan and James Thompson’s team at the University of Wisconsin) announced that they had successfully replicated the biological abilities of the embryonic stem cell using only skin cells. Called “induced pluripotent stem cells” these former skin cells were programmed to become other types of cells, acting in the same way as the embryonic stem cells.
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