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Autologous Stem Cell Therapy and its Effects on COPD: A Pilot Study

  |   COPD   |   No comment

Lung Institute, LLC

Author: Jack Coleman Jr., M.D.

 

Today more than 600 million people suffer from chronic obstructive pulmonary disease (COPD) worldwide. Although there is no cure for COPD, recent advancements in stem cell therapy have made it possible not only to relieve symptoms, but to promote healing within the lungs themselves.

 

After testing approximately 100 patients, the Lung Institute has discovered that within three months of treatment, 84 percent of patients found their quality of life had improved. Among other promising statistics this discovery could change lives significantly. For millions of people suffering from COPD, a natural decline in pulmonary health is a harsh reality. Based on these results, stem cell therapy could be the answer they’ve been looking for.

 

For more details and statistics on the effects of stem cell therapy on pulmonary function improvement, continue reading below.

 

The Problem with Chronic Pulmonary Diseases

 

Chronic Obstructive Pulmonary Disease (COPD) is a progressive lung disorder that often occurs as a result of prolonged cigarette smoking, second-hand smoke, and polluted air or working conditions. COPD is the most prevalent form of chronic lung disease. The physiological symptoms of COPD include shortness of breath (dyspnea), cough, and sputum production, exercise intolerance and reduced Quality of Life (QOL). These signs and symptoms are brought about by chronic inflammation of the airways, which restricts breathing. When fibrotic tissues contract, the lumen is narrowed, compromising lung function. As histological studies confirm, airway fibrosis and luminal narrowing are major features that lead to airflow limitation in COPD 1-3.

 

Today, COPD is a serious global health issue, with a prevalence of 9-10% of adults aged 40 and older4. And the prevalence of the disease is only expected to rise. Currently COPD accounts for 27% of tobacco related deaths and is anticipated to become the fourth leading cause of death worldwide by 2030 5. Today, COPD affects approximately 600 million individuals—roughly 5% of the world’s population 6. Despite modern medicine and technological advancements, there is no known cure for COPD.

 

The difficulty in treating COPD and other lung diseases rests in the trouble of stimulating alveolar wall formation15. Until recently, treatment has been limited by two things: a lack of understanding of the pathophysiology of these disease processes on a molecular level and a lack of pharmaceutical development that would affect these molecular mechanisms. This results in treatment focused primarily in addressing the symptoms of the disease rather than healing or slowing the progression of the disease itself.

 

The result is that there are few options available outside of bronchodilators and corticosteroids7. Although lung transplants are performed as an alternative option, there is currently a severe shortage of donor lungs, leaving many patients to die on waiting lists prior to transplantation. Lung transplantation is also a very invasive form of treatment, commonly offering poor results, a poor quality of life with a 5-year mortality rate of approximately 50%, and a litany of health problems associated with lifelong immunosuppression13.

 

However, it has been shown that undifferentiated multipotent endogenous tissue stem cells (cells that have been identified in nearly all tissues) may contribute to tissue maintenance and repair due to their inherent anti-inflammatory properties. Human mesenchymal stromal cells have been shown to produce large quantities of bioactive factors including cytokines and various growth factors which provide molecular cueing for regenerative pathways. This affects the status of responding cells intrinsic in the tissue 18. These bioactive factors have the ability to influence multiple immune effector functions including cell development, maturation, and allo-reactive T-cell responses 19. Although research on the use of autologous stem cells (both hematopoietic and mesenchymal) in regenerative stem cell therapy is still in the early stages of implementation, it has shown substantive progress in treating patients with few if any adverse effects.

 

Summary of Process
The Lung Institute (LI) provided treatment by harvesting autologous stem cells (hematopoietic stem cells and mesenchymal stromal cells) by withdrawing adipose tissue (fat), bone marrow or peripheral blood. These harvested cells are isolated and concentrated, and along with platelet-rich plasma, are then reintroduced into the body and enter the pulmonary vasculature (vessels of the lungs) where cells are trapped in the microcirculation (the “pulmonary trap”). Alternatively, nebulized stem cells are reintroduced through the airways in patients who have undergone an adipose (fat tissue) treatment.

 

Methodology
Individuals diagnosed with COPD were tracked by the Lung Institute to measure the effects of treatment via either the venous protocol or adipose protocol on both their pulmonary function as well as their Quality of Life.

 

Pulmonary Function Test (PFT)
All PFTs were performed according to national practice guideline standards for repeatability and acceptability8-10. On PFTs, pre-treatment data was collected through on-site testing or through previous medical examinations by the patient’s primary physician (if done within two weeks). The test was then repeated by their primary physician 6 months after treatment.*

* Due to the examination information required from primary physicians, only 25 out of 100 patients are reflected in the PFT data.

 

Quality of Life Survey (QLS) and Quality Improvement Score (QIS)
Patients with progressive COPD will typically experience a steady decrease in their Quality of Life. Given this development, a patient’s Quality of Life score is frequently used to define additional therapeutic effects, with regulatory authorities frequently encouraging their use as primary or secondary outcomes17.

 

On quality of life testing, data was collected through the implementation of the Clinical COPD Questionnaire (CCQ) based survey17. The survey measured the patient’s self-assessed quality of life on a 0-6 scale, with adverse Quality of Life correlated in ascending numerical order. It was implemented in three stages: pre-treatment, 3-months post-treatment, and 6-months post-treatment. The survey measured two distinct outcomes: the QLS score, which measured the patient’s self-assessed quality of life score, and the QIS, a percentage-based measurement determining the proportion of patients within the sample that experienced QLS score improvements.

patient-demographicsDemographics
Over the duration of six months, the results of 100 patients treated for COPD through venous and adipose based therapies were tracked by the Lung Institute in order to measure changes in pulmonary function and any improvement in Quality of Life.

 

Of the 100 patients treated by the Lung Institute, 64 were male (64%) and 36 were female (36%). Ages of those treated range from 55-88 years old with an average age of 71. Throughout the study, 82 (82%) were treated with venous derived stem cells, while 18 (18%) were treated from stem cells derived from adipose tissue.

 

 

 

copd-patients

* The survey measured the patient’s self-assessed quality of life on a 0-6 scale, with adverse Quality of Life correlated in ascending numerical order.

 

Over the course of the study, the patient group averaged an increase of 35.5% to their Quality of Life (QLS) score within three months of treatment. While in the QIS, 84% of all patients found that their Quality of Life score had improved within three months of treatment (figure 1.3).

quality-of-life-improvement

 

Within the PFT results, 48% of patients tested saw an increase of over 10% to their original pulmonary function with an average increase of 16%. During the three to six month period after treatment, patients saw a small decline in their progress, with QLS scores dropping from 35.5% to 32%, and the QIS from 84% to 77%.

 

Fletcher and Peto’s work shows that patient survival rate can be improved through appropriate or positive intervention14 (figure 1.4). It remains to be seen if better quality of life will translate to longevity, but if one examines what factors allow for improved quality of life such as improvement in oxygen use, exercise tolerance, medication use, visits to the hospital and reduction in disease flare ups then one can see that quality of life improves in association with clinical improvement.

 

 

lung-function-decline

Analysis
Currently the most utilized options for treating COPD are bronchodilator inhalers with or without corticosteroids and lung transplant – each has downsides. Inhalers are often used incorrectly11, are expensive over time, and can only provide temporary relief of symptoms. Corticosteroids, though useful, have risk of serious adverse side effects such as infections, blood sugar imbalance, and weight gain to name a few 16. Lung transplants are expensive, have an adverse impact on quality of life and have a high probability of rejection by the body the treatment of which creates a new set of problems for patients. In contrast, initial studies of stem cells treatments show efficacy, lack of adverse side effects and may be used safely in conjunction with other treatments.

 

 

Conclusion
Through the data collected by the Lung Institute, developing methodologies for this form of treatment are quickly taking place as other entities of the medical community follow suit. In a recent study of regenerative stem cell therapy done by the University of Utah, patients exhibited improvement in PFTs and oxygen requirement compared to the control group with no acute adverse events12. Through the infusion of stem cells derived from the patient’s own body, stem cell therapy minimizes the chance of rejection to the highest degree, promotes healing and can improve the patient’s pulmonary function and quality of life with no adverse side effects.

 

Although more studies using a greater number of patients is needed to further examine objective parameters such as PFTs, exercise tests, oxygen, medication use and hospital visits, larger sample sizes will also help determine if one protocol is more beneficial than others. With deeper research, utilizing economic analysis along with longer-term follow up will answer questions on patient selection, the benefits of repeated treatments, and a possible reduction in healthcare costs for COPD treatment.

 

The field of Cellular Therapy and Regenerative Medicine is rapidly advancing and providing effective treatments for diseases in many areas of medicine.The Lung Institutes strives to provide the latest in safe, effective therapy for chronic lung disease and maintain a leadership role in the clinical application of these technologies.

 

In a landscape of scarce options and rising costs, the Lung Institute believes that stem cell therapy is the future of treatment for those suffering from COPD and other lung diseases. Although data is limited at this stage, we are proud to champion this form of treatment while sharing our findings.
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Hogg J, Chu F, Utokaparch S, et al. The nature of small-airway obstruction in chronic obstructive pulmonary disease. N Engl J Med 2004; 350: 2645-53.
Niewoehner D. Structure-function relationships: the pathophysiology of airflow obstruction. In: Stockley R, Rnnard S. Rabe K, Celli B, eds. Chronic Obstructive Pulmonary Disease. Hoboken, NJ: Blackwell, 2007: 3-19.
Halbert R, Natoli J. Gano A, Badamgarav E, Buist A, Mannino D. Global burden of COPD: systematic review and meta-analysis. Eur Respir J 2006; 28: 423-32.
Fernando J Martinez, James F Donohue, Stephen I Rennard. The future of chronic obstructive pulmonary disease treatment-difficulties of and barriers to drug development. Lancet 2011; 378: 1027-37.
Michelle J. Hansen, Rosa C. Gualano, Steve Bozinovski, Ross Vlahos, Gary P. Anderson. Therapeutic prospects to treat skeletal muscle wasting in COPD (chronic obstructive lung disease). Pharmacology & Therapy 2006; 109: 162-172.
European Medicines Agency. Guideline on clinical investigation of medicinal products for the treatment of Chronic Obstructive Pulmonary Disease (COPD). London: European Medicines Agency, 2010.
Wanger J, Clausen JL, Coastes A, et al. Standardisation of the measurement of lung volumes. Eur Respir J 2005; 26(3): 511-22.
Miller MR, Hankinson J, Brusasco V, et al. Standardisation of spirometry. Eur Respir J 2005; 26(2):319-38. ATS/ERS statement on respiratory muscle testing. Am J Respir Crit Care Med 2002; 166(4):518-624.

 

Macintyre N, Crapo RO, Viegi G, et al. Standardisation of the single-breath determination of carbon monoxide uptake in the lung. Eur Respir J 2005; 26 (4): 720-35.

Int J Chron Obstruct Pulmon Dis. 2008 Sep; 3(3): 371–384.
J. Tuma, F. Silva, A.A. Winters, C. Bartlett, A.N. Patel. Maison de Sante. Lima, Peru; University of Utah, Salt Lake City, UT.

Viranuj Sueblinvong, MD and Daniel J. Weiss, M.D., PHD. Stem Cells and Cell Therapy Approaches in Lung Biology and Diseases. 2010.

Tantucci C, Modina D. Lung function decline in COPD. International Journal of Chronic Obstructive Pulmonary Disease. 2012; 7:95-99. doi:10.2147/COPD.S27480.

Massaro G, Massaro D. Retinoic acid treatment abrogates elastase-induced pulmonary emphysema in rats. Nas Med 1997; 3: 675-77.
Samy Suissa, Valerie Patenaude, Francesco Lapi, Pierre Ernst. Inhaled corticosteroids in COPD and the risk of serious pneumonia. Thorax 2013; 68: 1029-1036.

Clinical COPD Questionnaire. http://www.ccq.nl/. Accessed October 28, 2015.
Tracey L. Bonfield, Arnold I. Caplan. Adult Mesenchymal Stem Cells: An Innovative Therapeutic for Lung Diseases. Discovery Medicine. April 2010; 9(47): 337-345.

Giselle Chamberlain, James Fox, Brian Ashton, Jim Middleton. Mesenchymal Stem Cells: Their Phenotype, Differentiation Capacity, Immunological Features, and Potential for Homing. Stem Cells. 2007; 25:2739-2749.

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The Gene Hackers

  |   DNA Sequences   |   No comment

A powerful new technology enables us to manipulate our DNA more easily than ever before.

(FROM THE NEW YORKER – BY MICHAEL SPECKTOR ) NOVEMBER 16, 2015

At thirty-four, Feng Zhang is the youngest member of the core faculty at the Broad Institute of Harvard and M.I.T. He is also among the most accomplished. In 1999, while still a high-school student, in Des Moines, Zhang found a structural protein capable of preventing retroviruses like H.I.V. from infecting human cells. The project earned him third place in the Intel Science Talent Search, and he applied the fifty thousand dollars in prize money toward tuition at Harvard, where he studied chemistry and physics. By the time he received his doctorate, from Stanford, in 2009, he had shifted gears, helping to create optogenetics, a powerful new discipline that enables scientists to use light to study the behavior of individual neurons.

 

Zhang decided to become a biological engineer, forging tools to repair the broken genes that are responsible for many of humanity’s most intractable afflictions. The following year, he returned to Harvard, as a member of the Society of Fellows, and became the first scientist to use a modular set of proteins, called TALEs, to control the genes of a mammal. “Imagine being able to manipulate a specific region of DNA . . . almost as easily as correcting a typo,” one molecular biologist wrote, referring to TALEs, which stands for transcription activator-like effectors. He concluded that although such an advance “will probably never happen,” the new technology was as close as scientists might get.
Having already helped assemble two critical constituents of the genetic toolbox used in thousands of labs throughout the world, Zhang was invited, at the age of twenty-nine, to create his own research team at the Broad. One day soon after his arrival, he attended a meeting during which one of his colleagues mentioned that he had encountered a curious region of DNA in some bacteria he had been studying. He referred to it as a CRISPR sequence.
“I had never heard that word,” Zhang told me recently as we sat in his office, which looks out across the Charles River and Beacon Hill. Zhang has a perfectly round face, its shape accentuated by rectangular wire-rimmed glasses and a bowl cut. “So I went to Google just to see what was there,” he said. Zhang read every paper he could; five years later, he still seemed surprised by what he found. CRISPR, he learned, was a strange cluster of DNA sequences that could recognize invading viruses, deploy a special enzyme to chop them into pieces, and use the viral shards that remained to form a rudimentary immune system. The sequences, identical strings of nucleotides that could be read the same way backward and forward, looked like Morse code, a series of dashes punctuated by an occasional dot. The system had an awkward name—clustered regularly interspaced short palindromic repeats—but a memorable acronym.

 

CRISPR has two components. The first is essentially a cellular scalpel that cuts DNA. The other consists of RNA, the molecule most often used to transmit biological information throughout the genome. It serves as a guide, leading the scalpel on a search past thousands of genes until it finds and fixes itself to the precise string of nucleotides it needs to cut. It has been clear at least since Louis Pasteur did some of his earliest experiments into the germ theory of disease, in the nineteenth century, that the immune systems of humans and other vertebrates are capable of adapting to new threats. But few scientists had considered the possibility that single bacterial cells could defend themselves in the same way. The day after Zhang heard about CRISPR, he flew to Florida for a genetics conference. Rather than attend the meetings, however, he stayed in his hotel room and kept Googling. “I just sat there reading every paper on CRISPR I could find,” he said. “The more I read, the harder it was to contain my excitement.”
It didn’t take Zhang or other scientists long to realize that, if nature could turn these molecules into the genetic equivalent of a global positioning system, so could we. Researchers soon learned how to create synthetic versions of the RNA guides and program them to deliver their cargo to virtually any cell. Once the enzyme locks onto the matching DNA sequence, it can cut and paste nucleotides with the precision we have come to expect from the search-and-replace function of a word processor. “This was a finding of mind-boggling importance,” Zhang told me. “And it set off a cascade of experiments that have transformed genetic research.”

 

With CRISPR, scientists can change, delete, and replace genes in any animal, including us. Working mostly with mice, researchers have already deployed the tool to correct the genetic errors responsible for sickle-cell anemia, muscular dystrophy, and the fundamental defect associated with cystic fibrosis. One group has replaced a mutation that causes cataracts; another has destroyed receptors that H.I.V. uses to infiltrate our immune system.

 

The potential impact of CRISPR on the biosphere is equally profound. Last year, by deleting all three copies of a single wheat gene, a team led by the Chinese geneticist Gao Caixia created a strain that is fully resistant to powdery mildew, one of the world’s most pervasive blights. In September, Japanese scientists used the technique to prolong the life of tomatoes by turning off genes that control how quickly they ripen. Agricultural researchers hope that such an approach to enhancing crops will prove far less controversial than using genetically modified organisms, a process that requires technicians to introduce foreign DNA into the genes of many of the foods we eat.

 

The technology has also made it possible to study complicated illnesses in an entirely new way. A few well-known disorders, such as Huntington’s disease and sickle-cell anemia, are caused by defects in a single gene. But most devastating illnesses, among them diabetes, autism, Alzheimer’s, and cancer, are almost always the result of a constantly shifting dynamic that can include hundreds of genes. The best way to understand those connections has been to test them in animal models, a process of trial and error that can take years. CRISPR promises to make that process easier, more accurate, and exponentially faster.

 

Inevitably, the technology will also permit scientists to correct genetic flaws in human embryos. Any such change, though, would infiltrate the entire genome and eventually be passed down to children, grandchildren, great-grandchildren, and every subsequent generation. That raises the possibility, more realistically than ever before, that scientists will be able to rewrite the fundamental code of life, with consequences for future generations that we may never be able to anticipate. Vague fears of a dystopian world, full of manufactured humans, long ago became a standard part of any debate about scientific progress. Yet not since J. Robert Oppenheimer realized that the atomic bomb he built to protect the world might actually destroy it have the scientists responsible for a discovery been so leery of using it.

 

To read the full article, click here.

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Scientists time-reverse developed stem cells to make them ‘embryonic’ again

  |   Embroyonic Stem Cells   |   No comment

May help avoid ethically controversial use of human embryos for research and support other research goals.

From KURZWEILAI 

 

University of Michigan Medical School researchers have discovered a way to convert mouse stem cells (taken from an embryo) that have  become “primed” (reached the stage where they can differentiate, or develop into every specialized cell in the body) to a “naïve” (unspecialized) state by simply adding a drug.

 

This breakthrough has the potential to one day allow researchers to avoid the ethically controversial use of human embryos left over from infertility treatments. To achieve this breakthrough, the researchers treated the primed embryonic stem cells (“EpiSC”) with a drug called MM-401* (a leukemia drug) for a short period of time.

 

Reverting back to the embryonic state

As the research team reports in the journal Cell Stem Cell, this drug treatment caused more than half of these EpiSC cells to return to a naïve (less specialized) state as “reverted embryonic stem cells” (rESC). The researchers then bred healthy mice from those reverted rESC cells — proving that the drug-treated cells were still viable and had the ability to become any type of cell (achieved pluripotency).

 

This study is significant because it’s the first time scientists have been able to make stem cells revert to their original state without complications. Also, the drug leaves no trace behind, whereas genetic modification of the stem cells (used by other researchers) may block the stem cells from developing into healthy cells. The researchers only needed to treat the EpiSCs with a single drug and for just a few days.

 

Past attempts by other teams to return mouse EpiSC cells to the original naïve state have either resulted in a far lower proportion of cells returned to a reverted state, or have produced cells that were not viable. Those past studies also needed to use cocktails of multiple drugs given over the long term.

 

The work was funded by the National Institutes of Health and the Leukemia and Lymphoma Society.

 

Human stem cells next

Meanwhile, scientists at the University of Cambridge have also just reported (in an open-access paper in the journal Stem Cell Reports) that they have also produced stem cells directly from embryos — but they did it with human stem cells (for the first time).

 

“Until now it hasn’t been possible to isolate these naïve stem cells, even though we’ve had the technology to do it in mice for thirty years — leading some people to doubt it would be possible,” says Ge Guo, the study’s first author and research associate in the Stem Cell Potency group at Cambridge.

 

“But we’ve managed to extract the cells and grow them individually in culture. Naïve stem cells have many potential applications, from regenerative medicine to modeling human disorders.”

 

Jenny Nichols, PhD,  joint senior author of the study, says that one of the most exciting applications of their new technique would be to study disorders that arise from cells that contain an abnormal number of chromosomes. Ordinarily, the body contains 23 pairs of identical chromosomes (22 pairs and one pair of sex chromosomes), but some children are born with additional copies, which can cause problems. For example, children with Down’s syndrome are born with three copies of chromosome 21.

 

“Even in many ‘normal’ early-stage embryos, we find several cells with an abnormal number of chromosomes,” explains Nichols. “Because we can separate the cells and culture them individually, we could potentially generate ‘healthy’ and ‘affected’ cell lines. This would allow us to generate and compare tissues of two models, one ‘healthy’ and one that is genetically-identical other than the surplus chromosome. This could provide new insights into conditions such as Down’s syndrome.”

 

The research was supported by the Medical Research Council, Biotechnology and Biological Sciences Research Council, Swiss National Science Foundation, and the Wellcome Trust.

 

* The drug, MM-401, specifically targets epigenetic chemical markers on histones, the protein “spools” that DNA coils around to create structures called chromatin. These epigenetic changes signal the cell’s DNA-reading machinery and tell it where to start uncoiling the chromatin in order to read it.

A gene called Mll1 is responsible for the addition of these epigenetic changes, which are like small chemical tags called methyl groups. Mll1 plays a key role in the uncontrolled explosion of white blood cells in leukemia, which is why researchers developed the drug MM-401 to interfere with this process. But Mll1 also plays a role in cell development and the formation of blood cells and other cells in later-stage embryos.

Stem cells do not turn on the Mll1 gene until they are more developed. The MM-401 drug blocks Mll1’s normal activity in developing cells so the epigenetic chemical markers are missing. These cells are then unable to continue to develop into different types of specialized cells but are still able to revert to healthy naive pluripotent stem cells.

 

Abstract of MLL1 Inhibition Reprograms Epiblast Stem Cells to Naive Pluripotency

The interconversion between naive and primed pluripotent states is accompanied by drastic epigenetic rearrangements. However, it is unclear whether intrinsic epigenetic events can drive reprogramming to naive pluripotency or if distinct chromatin states are instead simply a reflection of discrete pluripotent states. Here, we show that blocking histone H3K4 methyltransferase MLL1 activity with the small-molecule inhibitor MM-401 reprograms mouse epiblast stem cells (EpiSCs) to naive pluripotency. This reversion is highly efficient and synchronized, with more than 50% of treated EpiSCs exhibiting features of naive embryonic stem cells (ESCs) within 3 days. Reverted ESCs reactivate the silenced X chromosome and contribute to embryos following blastocyst injection, generating germline-competent chimeras. Importantly, blocking MLL1 leads to global redistribution of H3K4me1 at enhancers and represses lineage determinant factors and EpiSC markers, which indirectly regulate ESC transcription circuitry. These findings show that discrete perturbation of H3K4 methylation is sufficient to drive reprogramming to naive pluripotency.

 

Abstract of Naive Pluripotent Stem Cells Derived Directly from Isolated Cells of the Human Inner Cell Mass

Conventional generation of stem cells from human blastocysts produces a developmentally advanced, or primed, stage of pluripotency. In vitro resetting to a more naive phenotype has been reported. However, whether the reset culture conditions of selective kinase inhibition can enable capture of naive epiblast cells directly from the embryo has not been determined. Here, we show that in these specific conditions individual inner cell mass cells grow into colonies that may then be expanded over multiple passages while retaining a diploid karyotype and naive properties. The cells express hallmark naive pluripotency factors and additionally display features of mitochondrial respiration, global gene expression, and genome-wide hypomethylation distinct from primed cells. They transition through primed pluripotency into somatic lineage differentiation. Collectively these attributes suggest classification as human naive embryonic stem cells. Human counterparts of canonical mouse embryonic stem cells would argue for conservation in the phased progression of pluripotency in mammals.

 

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