Patient’s Own Stem Cells Treat Rare Neurological Disorder

Stiff-Person syndrome is a rare neurological disease that, for all intents and purposes, looks like an autoimmune disease. It is characterized by muscular rigidity that tends to come and go. This rigidity occurs in the muscles of the trunks and limbs. Patients with Stiff-Person syndrome also have an enhanced sensitivity to stimuli such as noise, touch, and emotional distress, and various stimuli may cause the patient to experience painful muscle spasms that cause abnormal postures and stiffening. Stiff-Person syndrome or SPS is more common in women than in men and SPS patients often suffer from other autoimmune conditions in addition to SPS (for example, pernicious anemia, diabetes, vitiligo, and thyroiditis). Unfortunately, the precise cause of SPS is not known, but again, it looks like an autoimmune condition.

A research team at Ottawa Hospital Research Institute has made a breakthrough in the successful treatment of SPS using bone marrow stem cell transplants. The medical director at the Ottawa Hospital Research Institute, Dr. Harold L. Atkins, who is also a physician in the Blood and Bone Marrow Transplant Program at The Ottawa Hospital and an associate professor at the University of Ottawa has used bone marrow transplants to two female SPS patients into remission.

SPS can leave patients bedridden and in severe pain, but thanks to Atkins and his team, the progression of the disease in these women has ceased, allowing both women to regain their previous function and leaving them well enough to return to work and normal everyday activities.

Adkins and his group published this case study in JAMA Neurology, which is produced by the Journal of the American Medical Association. This is the first documented report that taking stem cells from a person’s own body can produce long-lasting remission of stiff person syndrome.

“We approach these cases very carefully and are always aware that there have just been a few patients treated and followed for a short time,” says Dr. Atkins. Atkins and his extracted bone marrow stem cells from each woman, and then used chemotherapy to eliminate their immune systems. Once their immune system were reliably eliminated, both women had their own stem cells returned to their bodies in order to reconstitute their immune systems. This procedure essentially gives the immune system a “do-over.”.

“By changing the immune system, one hopes to put the stiff person syndrome into remission,” adds Dr. Atkins. “Seeing these two patients return to their normal lives is really every physicians dream.”

This very procedure, which is known as an “autologous stem cell transfer” or ASCT has been used to successfully treat people who suffer from autoimmune diseases such as multiple sclerosis, scleroderma, and systemic lupus erythematosis. Atkins and his team used high-doses of chemotherapy and antibodies that specifically bind lymphocytes to rid the women’s bodies of their rogue immune cells before their immune systems were regenerated using their own stem cells. Adkins an his colleagues viewed this as a viable treatment option based strategies that had been used to treat other autoimmune diseases.

Patient 1 was diagnosed with stiff person syndrome in 2005 at age 48 after experiencing leg stiffness and several falls. After her treatment, her symptoms disappeared and she was fully mobile again six months after receiving the stem cell transplant procedure in 2009.

Patient 2 was diagnosed with stiff person syndrome in 2008 at age 30. She had stopped working and driving, and had moved back in with her parents before her stem cell transplant in 2011. Also, she has been able to return to her work and previous activities, and has not had any stiff person syndrome symptoms in more than a year.

“The results achieved by Dr. Atkins and his team through this innovative treatment show how research at The Ottawa Hospital can lead to life-changing and, even life-saving care,” says Dr. Duncan Stewart, Chief Executive Officer and Scientific Director of the Ottawa Hospital Research Institute. “Translating research into better care for patients is what we’re all about at the research institute.”

A More Efficient Way to Make Induced Pluripotent Stam cells

Mark Stadtfeld and his colleagues at the NYU Longone Medical Center has devised a new method for making induced pluripotent stem cells that greatly increases efficiency at which these cells are made.

Induced pluripotent stem cells or iPSCs are made from mature, adult cells by mean of a combination of genetic engineering and cell culture techniques. In short, the expression of four genes is forced in adult cells; Oct4, Sox2, Klf4, and c-Myc or OSKM. The proteins encoded by these four genes cooperatively work to drive a fraction of the cells into an immature state that resembles that of embryonic stem cells. These cells are them grown in cell culture systems that select for those cells that can grow continuously and form colonies of cells derived from progenitor cells. These cell colonies are them repeated isolated a re-cultured until an iPSC line has been established.

Unfortunately, this process is rather inefficient and tedious, since less than one percent or so of the reprogrammed cells actually undergo successful reprogramming. Additionally, it can take several weeks to properly establish an iPSC line. Thus, stem cell scientists have been looking at several different ways to boost the efficiency of this process.

Stadtfeld and his coworkers tried to add compounds to the cultured cells to determine if the culture conditions could actually augment the efficiency of the reprogramming process. “We especially wanted to know if these compounds could be combined to obtain stem cells at high-efficiency,” said Stadtfeld.

The compounds to which Stadtfeld was referring were two cell signaling proteins called Wnt and TFG-beta. Both of these compounds regulate a host of cell growth processes. Stadtfeld wanted to try regulating both of these pathways at the same time, in addition to providing cells with ascorbic acid, which is also known as vitamin C. Even vitamin C is more popularly known as an antioxidant, vitamin C also can remodel chromatin (that tight structure into which cells package their DNA).

When mouse skin fibroblasts were treated with OSKM and a compound that activates Wnt signaling, the efficiency of iPSC derivation increased slightly. The same thing was observed if fibroblasts were treated with OSKM and a compound that inhibits TGF-beta signaling or vitamin C. However, when all three of these compounds were combined, OSKM-engineered fibroblasts were reprogrammed at an efficiency of close to 80 percent. When different cell types were used as the starting cell, such as blood progenitor cells, the efficiency jumped to close to 100 percent; a result that was also observed if liver progenitor cells were used as the starting cell.

Stadtfeld is confident that these dramatic increases in iPSC derivation should improve future studies with iPSCs, since his protocol should make iPSC derivation more predictable. “It’s just a lot easier this way to study the mechanisms that govern reprogramming, as well as detect any undesired features that might develop in iPSCs,” he said.

Vitamin C and the two compounds used to manipulate the Wnt and TGF-β pathways have been widely used in research and have few unknown or hazardous effects. However, OKSM has in some cases caused undesired features in iPSCs, such as increased mutation rates. Stadtfeld believes that by making iPSC induction more rapid and efficient, his new technique might also make the resulting stem cells safer. “Conceivably it reduces the risk of abnormalities by smoothening out the reprogramming process,” Dr. Stadtfeld says. “That’s one of the issues we’re following up.”

Embryonic Stem Cell-Derived Retinal Cells Treat Blindness in Eye Patients

Embryonic stem cells are derived from human embryos, can only grow in culture indefinitely, and have the ability to potentially differentiate into any adult cell type in the human body.  Because cell and tissues made from embryonic stem cells bear the same tissue types as the embryos from which they were derived, they will be rejected by the immune system patient.  However, there are sites in our bodies were the immune system does not go, and that includes the central nervous system and the eyes.  This is the reason why clinical trials with embryonic stem cell-derived cells have focused, to date, on spinal cord injuries and eye diseases.

Several clinical trials have examined the ability of retinal pigmented epithelial (RPE) cells made from embryonic stem cells to treat patients with dry macular degeneration or an inherited eye disease called Stargardt’s disease.  Data from these trials has been reported in an article in the medical journal The Lancet, and accordingly, none of the treated patients showed tumor formation or immunological rejection of the implants and, most impressively perhaps, partial blindness was reversed in about half of the eyes that received transplants.

The results might re-energize the quest to harness embryonic stem cells for human medicine.  Dr. Anthony Atala of the Wake Forest Institute for Regenerative Medicine called the work “a major accomplishment” in an accompanying commentary on the article.

RPE cells lie just behind the photoreceptor cells in the retina of our eyes.  Photoreceptors have their ends hurried in the RPE layer.  This arrangement exists for a very good reason; the photoreceptors are exposed to high intensities of light and they suffer respectable amounts of oxidative damage.  The components of the photoreceptors cells are made in the very lowest parts of the RPEs and then are eventually pushed to the ends of the cells.  At the end of the photoreceptor cells, the RPEs relieve the photoreceptors of their photodamaged parts and gobble them down, and recycle the cellular components.  Thus, RPE cells serve a photoreceptor cell repair and service cells.  If the RPE cells begin to die, the photoreceptors are not long the this work either.

In the case of dry macular degeneration, which accounts for 90 percent of diagnosed cases of macular degeneration, the light-sensitive photoreceptor cells of the macula (the portion of the retina were the day vision is the sharpest) slowly break down. Damage to the macula causes blurring or spotty loss of central vision and yellowish cellular deposits called drusen (extracellular waste products from metabolism) form under the retina between the retinal pigmented epithelium (RPE) layer and a basement membrane called Bruch’s membrane, that supports the retina. An increase in the size and number of drusen is associated with the death of RPE and, consequently, photoreceptor cells, and is sometimes the first sign of dry macular degeneration.

Medical illustration of dry macular degeneration

Mutations in several genes have been identified in families with dry macular degeneration that increase the risk for dry macular degeneration.  These include the SERPING1 gene, those genes that encode the complement system proteins  factor H (CFH), factor B (CFB) and factor 3, and fibulin-5.  Additionally, some environmental and behavioral factors also influence the risk a person will develop macular degeneration.  These include smoking, exposure to blue light, ingestion of a high-fat diet, elevated blood pressure and serum cholesterol levels, and low vitamin D levels.

Stargardt’s disease is an inherited, juvenile form of macular degeneration that is caused by mutations in the ABCR gene.  The protein encoded by this gene is a waste metabolite transporter, and defects in this protein cause the build up of a toxic metabolite called lipofuscin in the RPE cells, which leads to their demise and the death of the photoreceptors.

In this study, the main goal was to assess the safety of the transplanted cells. The study “provides the first evidence, in humans with any disease, of the long-term safety and possible biologic activity” of cells derived from embryos, said co-author Dr. Robert Lanza, chief scientific officer of Advanced Cell Technology, which produced the cells and funded the study.

Nine patients with Stargardt’s disease and nine with dry age-related macular degeneration received implants of the retinal cells in one eye. The other eye served as a control.  Four eyes developed cataracts and two became inflamed, probably due to the patients’ age (median: 77) or the use of immune-supressing transplant drugs.

The implanted RPE cells survived in all 18 patients, most of whose vision improved.  In those with macular degeneration, treated eyes saw a median of 14 additional letters on a standard eye chart a year after receiving the cells, with one patient gaining 19 letters. The untreated eyes got worse, overall. The Stargardt’s patients had similar results.

In real-life terms, patients who couldn’t see objects under 12 feet (4 meters) tall can now see normal-size adults.

The vision of one 75-year old rancher who was blind in the treated eye (20/400) improved to 20/40, enough to ride horses again, Lanza said.  Others became able to use computers, read watches, go to the mall or travel to the airport alone for the first time in years.

While calling the results “encouraging,” stem cell expert Dusko Ilic of Kings College London, who was not involved in the work, warned that even if the larger clinical trial planned for later this year is also successful, “it will take years before the treatment becomes available.”

Other cell types can also form RPE cells and these include induced pluripotent stem cells, mesenchymal stem cells from fat (Ophthalmic Res. 2012;48 Suppl 1:1-5), adult retinal stem cells (Pigment Cell Melanoma Res. 2011 Feb;24(1):233-40), and iris pigmented epithelial cells (Prog Retin Eye Res. 2007 May;26(3):302-21).  We do not need to destroy embryos to treat eye diseases with stem cells.

Nose Stem Cells Help Bulgarian Man Walk With Braces

Darek Fidyka, a 38-year-old Bulgarian man, was severely injured by a stab wound in 2010 and consequently lost the ability to walk.

Now, a new procedure using stem cells from his nose has given him the ability to walk with the help of braces.

Olfactory ensheathing cells or OECs (also known as olfactory ensheathing glial or OEGs) are found in the olfactory system, inside the skull and in the covering of cells that lines the roof of the nose. OECs share similarities to other glial cells like Schwann cells, astrocytes, and oligodendrocytes. OECs can aid the extension of neural projections known as axons from the nasal tissue to the olfactory glomeruli. OECs can do this because they secrete several interesting neurotrophic factors and cell adhesion molecules and migrate along with the regenerating axons. Because of these properties, OECs can escort axonal extension through glial scars that are made in a spinal cord after a spinal cord injury. These scars inhibit the outgrowth of new axons but OECs can allow regenerating axons to bridge these glial scars.

An advantage of OECs is that they can coexist with astrocytes, the cells that contribute to the formation of the glial scar, and even seem to prevent the out-of-hand response astrocytes have in response to injury in which they synthesize a host of molecules that inhibit axon regeneration called “inhibitory proteoglycans.”

The pioneering technique used in this procedure, according to Geoffrey Raisman, a professor at University College London’s (UCL) institute of neurology, used OECs to construct a kind of bridge between two stumps of the damaged spinal column.

“We believe… this procedure is the breakthrough which, as it is further developed, will result in a historic change in the currently hopeless outlook for people disabled by spinal cord injury,” said Riesman, who led this research project.

Raisman, who is a spinal injury specialist at UCL, collaborated with neurosurgeons at Wroclaw University Hospital in Poland to remove one of Fidyka’s olfactory bulbs, which give people their sense of smell, and transplant his olfactory ensheathing cells (OECs) in combination with. olfactory nerve fibroblasts (ONFs) into the damaged spinal cord areas. Following 19 months of treatment, Fidyka recovered some voluntary movement and some sensation in his legs.

The Nicholls Spinal Injury Foundation, a British-based charity which part-funded the research, said in statement that Fidyka was continuing to improve more than predicted, and was now able to drive and live more independently.

OECs have been used before to treat spinal cord injury patients. I refer you to chapter 27 in my book, The Stem Cell Epistles, to learn more about these. The novel technique in this paper is the additional use of nasal fibroblasts and the construction of a bridge between the two damaged remnants of the spinal cord.

The reason OECs were recruited to treat spinal cord injuries is that when axons that carry information about smells are damaged, the neuron simply regenerates its atonal extension, which grows into the olfactory bulbs. OECs facilitate this process by re-opening the surface of the olfactory bulbs in order for the new axons to enter them. Thus Raisman and others have the notion that transplanted OECs in the damaged spinal cord could equally facilitate the regeneration of severed nerve fibers.

Raisman also added that the technique used in this case, that is bridging the spinal cord with nerve grafts from the patient, had been used in animal studies for years, but was never used in a human patient in combination with OECs.

“The OECs and the ONFs appeared to work together, but the mechanism between their interaction is still unclear,” he said in a statement about the work.

Several spinal cord injury experts who were not directly involved in this work said its results offered some new hope. However, they were also quick to add that more work needed to be done to precisely determine what had led to this success. More patients must be successfully treated with this procedure before its potential can be properly assessed.

“While this study is only in one patient, it provides hope of a possible treatment for restoration of some function in individuals with complete spinal cord injury,” said John Sladek, a professor of neurology and pediatrics at the University of Colorado School of Medicine in the United States.

Raisman and his team now plan to repeat the treatment technique in between three and five spinal cord injury patients over the next three to five years. “This Nose will enable a gradual optimization of the procedures,” he told Reuters.

Stem Cell Transplant from Gut Repairs Damaged Gut in Mice with Inflammatory Bowel Disease

Even though a stem population has been identified and studied in the gastrointestinal tract, Wellcome Trust Researchers have identified a new source of GI-based stem cells that have the ability to repair damage from inflammatory bowel disease when transplanted into mice.  This work comes to us from the Wellcome Trust-Medical Research Council Cambridge Stem Cell Institute at the University of Cambridge and BRIC at the University of Copenhagen, Denmark.  This work could translate into patient-specific regenerative therapies for inflammatory bowel diseases such as ulcerative colitis.

Adult tissues contain specialized stem cells populations that maintain individual tissues and organs.  Adult stem cells tend to be restricted to their tissue of origin and also tend to have the ability to differentiate into a limited subset of adult cell types.  Stem cells found in the gut, for example, typically can typically contribute to the replenishment of the gut whereas stem cells in the skin will only contribute to maintenance of the skin.

When examining the developing intestinal tissue in a mouse embryos, Kim Jensen and her team discovered stem cell population hat were quite different from those adult stem cells that have been described in the gut.  These cells actively divided and also could be grown in the laboratory over long periods of time without undergoing differentiation into mature cells.  Under specific culture conditions, however, these cells could be induced to differentiate into mature intestinal tissue.

When these cells were transplanted into mice that suffered from an inflammatory bowel disease, The implanted stem cell attached to the damaged areas within the intestine, and began to integrate into the existing tissue, within three hours of implantation.

The lead researcher in this study, Dr. Kim Jensen, a Wellcome Trust researcher and Lundbeck foundation fellow, said: “We found that the cells formed a living plaster over the damaged gut. They seemed to respond to the environment they had been placed in and matured accordingly to repair the damage.

“One of the risks of stem cell transplants like this is that the cells will continue to expand and form a tumour, but we didn’t see any evidence of that with this immature stem cell population from the gut.”

Cells with similar characteristics were isolated from both mice and humans.  Jensen’s team also generated similar cells by reprogramming adult human cells to make induced Pluripotent Stem Cells (iPSCs) that were also grown under the appropriate culture conditions.

“We’ve identified a source of gut stem cells that can be easily expanded in the laboratory, which could have huge implications for treating human inflammatory bowel diseases. The next step will be to see whether the human cells behave in the same way in the mouse transplant system and then we can consider investigating their use in patients,” added Dr Jensen.

Human Umbilical Mesenchymal Stem Cells Decreases Dextran Sulfate Sodium-Induced Colitis in Mice

Ulcerative colitis is one of the Inflammatory Bowel Diseases (IBDs) that features chronic inflammation of the large intestine. This is an autoimmune disease that features constant attacks by the immune system on the intestinal mucosae, and the inner layer of the large intestine undergoes constant damage and healing, which increases the risk of the patient to developing colorectal carcinoma.

Mesenchymal stem cells have the capability to suppress inflammation, which makes them promising tools for treating diseases like ulcerative colitis. Unfortunately, the lack of reproducible techniques for harvesting and expanding MSCs has prevented bone marrow- and umbilical cord blood-derived MSCs from being routinely used in clinical situations.

However, a study that was published in the journal Clinical and Experimental Pharmacology and Physiology has used Wharton’s jelly derived umbilical MSCs (UMSCs) to treat mice in which an experimental form of ulcerative colitis was induced. Dextran sulfate sodium (DSS) induced colitis in mice has many of the pathological features of ulcerative colitis in humans.

When mice treated with DSS were also given Wharton’s jelly derived UMSCs showed significant diminution of the severity of colitis. The structure of the tissue in the colon looked far more normal and the types of molecules produced by inflammation were significantly reduced. In addition, transplantation of UMSCs reduced the permeability of the intestine and also increased the expression of tight junction proteins, which help knit the colonic cells together and maintain the structural integrity of the colon. These results show that the anti-inflammatory properties of UMSCs and their capacity to regulate tight junction proteins ameliorates ulcerative colitis.

U of Pitt Team Discovers Stem Cells in the Esophagus

Even though several studies have been unsuccessful at identifying a stem population in the esophagus, a study from the University of Pittsburgh has discovered a stem cell pool that services the esophagus. Researchers from the University of Pittsburgh School of Medicine have published an animal report in the journal Cell Reports that might lead to new insights into the development and treatment of esophageal cancer and a precancerous condition known as Barrett’s esophagus.

In the US, more than 18,000 people will be diagnosed with esophageal cancer in 2014 and almost 15,500 people will die from it, according to numbers generated by the American Cancer Society. The precancerous condition known as Barrett’s esophagus is characterized by tissue changes in the lining of the esophagus in which the esophageal lining begins to resemble the tissue architecture of the intestine. Barrett’s esophagus is usually a long-term consequence of gastro-esophageal reflux disease or GERD.

“The esophageal lining must renew regularly as cells slough off into the gastrointestinal tract,” said senior investigator Eric Lagasse, Pharm.D., Ph.D., associate professor of pathology, Pitt School of Medicine, and director of the Cancer Stem Cell Center at the McGowan Institute for Regenerative Medicine. “To do that, cells in the deeper layers of the esophagus divide about twice a week to produce daughter cells that become the specialized cells of the lining. Until now, we haven’t been able to determine whether all the cells in the deeper layers are the same or if there is a subpopulation of stem cells there.”

Lagasse and his team grew small explants of esophageal tissue in culture. These esophageal “organoids” from mice were then used to conduct experiments that were used to identify and track the different cells in the basal layer of the tissue. In these organoids, Lagasse and others found a small population of cells that divide more slowly, are less mature, can differentiate into several different types of esophageal-specific cell types, and have the ability to self-renew. The ability to self-renew is a defining feature of stem cells.

“It was thought that there were no stem cells in the esophagus because all the cells were dividing rather than resting or quiescent, which is more typical of stem cells,” Dr. Lagasse noted. “Our findings reveal that there indeed are esophageal stem cells, and rather than being quiescent, they divide slowly compared to the rest of the deeper layer cells.”

Lagasse and his team would now like to examine human esophageal tissues from patients with Barrett’s esophagus in order to determine if such patients show evidence of esophageal stem cell dysfunction.

“Some scientists have speculated that abnormalities of esophageal stem cells could be the origin of the tissue changes that occur in Barrett’s disease,” Dr. Lagasse said. “Our current and future studies could make it possible to test this long-standing hypothesis.”