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"Off-the-shelf" Engineered Stem Cells: Are They Therapeutically Valuable?

FROM THE DIRECTOR The answer is yes. But painfully slowly for Bedford Research Foundation scientists are the patients and families in need. Why? pursuing a promising example of The astounding 2013 report by an Oregon engineering stem cells -- to have a research team of the successful creation of Because new therapies involve both new resistance to HIV infection. The work stem cells from a somatic cell nucleus scientific discoveries and additional tests follows a proof-of-principle report by a transferred into an unfertilized human egg for medical safety. Demonstrating that German medical team in 2009. Because was met with surprising calm by the lay stem cells can turn into heart muscle or HIV infects the immune system, it is press and the bioethics community. This is nerve cells in a petri dish is exciting, but theoretically treatable by bone marrow in sharp contrast to the outcry a decade very far from therapeutic use. stem cell transplant. But past attempts ago when similar experiments were have shown transplanted bone marrow denounced as “human cloning” and the One big stumbling block is the source of becomes HIV infected, making it an U.S. congress rumbled with attempts to the stem cells to be used for therapies. unsuitable treatment approach for HIV outlaw all such research with human eggs. Must they be patient-specific? Or could a disease. This changed when the bone Public concern was further fueled by an bank of fully characterized, "off-the-shelf” marrow transplant in Germany resulted in extraordinary scientific fraud in 2005 by a stem cells be created? Stem cells that could an apparent cure of the patient’s HIV South Korean research team that falsely be administered immediately in the disease -- because the transplanted bone claimed to have created such stem cells. emergency room when the heart attack, stroke or spinal cord injury occurs? marrow stem cells were naturally deficient The 2013 calm is a clear, positive sign that Perhaps while patient-specific stem cells in CCR5, an receptor on the surface of cells the newness -- and the shock -- of the were being created? important for HIV infection. promise of stem cell regenerative medicine has worn off. Thousands of young According to estimates, only a scientists have been trained since the 1999 few hundred stem cell lines cover of Science announced stem cells as could tissue match more than the “breakthrough of the year.” 95% of the world’s population. So where are the stem cell therapies? This is an exciting prospect and an Where are the cures for diabetes, spinal achievable goal. The stem cells could even cord injuries/diseases, stroke, HIV/AIDS, be genetically engineered if needed to treat Parkinson’s disease, heart and kidney specific conditions, such as HIV/AIDS. failure? Are they coming? Continued on page 4

MORE NEWS INSIDE FALL 2013 - WINTER 2014 Prostate Disease Discovering Key Testis Project Update New Staff At The Donate to the Research Update Differences in Mouse Thanks to private Foundation Foundation Developing a better Embryos and donations, scientists are Alexis Agnew and Valia Your private donations screening test for Parthenotes in Phase 3 of the testis Dinopoulou join the make our important prostate cancer. Circadian cycles at work. stem cell project. BSCRF team. work possible. BSCRF Scientists Discover Developmental Differences Between Mouse Embryos and Parthenotes

The current goal of BSCRF research is to 80 Fertilized optimize the efficiency of deriving pluripotent 70 2 cell stem cells from testis and unfertilized human 3 Cell 60 39 eggs (parthenotes) for patient specific and 4 Cell 50 5 Cell 38 perhaps stem cell bank use. 6 cell 40 37 7 Cell In 2009, Bedford Research scientists 30 8 Cell discovered that circadian rhythm genes are Number of Embryos 36 Blastocyst 20 “on” in early human embryos, suggesting 10 35 circadian signals may be important to stem 0 34 cell derivation and stability. If true, new 12 16 20 24 28 32 36 40 44 48 52 56 60 64 68 72 76 80 84 88 92 96 100 106 methods of culturing stem cells in hours post-Fertilization laboratories that mimic circadian signals 60 Parthenotes 39 need to be developed. 50 38 40 In the body, the rhythm of circadian genes 37 30 is supported by several types of signals, 36 including light/dark cycles, hormone 20 Number of Embryos 35 pulses, body temperature variations, and 10 eating. The signals regulate the pattern of 0 34 circadian genes turning “on” and “off” in 13 17 21 25 29 33 37 41 45 49 53 57 61 65 69 73 77 81 85 89 93 97 103 109 115 121 127 133 hours post-Activation 24 hour cycles. In contrast, to date, stem cells have been cultured in constant 24 28 32 36 40 44 48 52 56 60 64 68 72 76 80 84 88 92 96 102 108 114 120 126 132 138 144 temperature in the dark, their only hours post-hCG potential circadian signal being renewal of their culture medium. Figure 2: Histogram of timing of cell cleavages following fertilization (top) or parthenogenetic activation (bottom). Wavy background line is temperature (right axis). To begin to understand the importance of circadian temperature oscillations to stem indicate the first cleavage of a mouse egg to two-cells takes place at approximately the cell derivation and expansion, BSCRF same time after fertilization or parthenogenic activation, the second cleavage to 3 cells scientists have taken advantage of their and 4 cells is markedly delayed in the parthenotes, the intervals to the third cleavages (5 newly developed “circadian incubator time to 8 cells) are approximately the same, but development to blastocyst is again delayed in lapse videomicroscope” to chronicle the the parthenotes. This indicates the parthenotes need additional developmental support first five days of development of mouse at the 2-cell stage and at the 8- to 16-cell stage. Discovering the needed support, and its embryos and parthenotes, which are being relationship to circadian signals, may markedly improve testis and parthenote stem cell used as models because cell division is derivation, and speed up the project to derive genetically modified parthenote stem cells. easy to see in a group of mouse embryos/ parthenotes. The goal is to discover if temperature oscillations play an important Learn More And Donate Now role in stem cell derivation or Scan this bar with your smart phone to watch differentiation into useful cell types, such as neurons or bone marrow stem cells. time lapse videos, learn more about on going The work is ongoing. research, and donate instantly.

As shown in Figure 2, results to date www.bedfordresearch.org 2 Progress In At The ISSCR Prostate Disease Testis Stem Cells In June, 2013, BSCRF presented a poster Research Update and hosted a booth at the 11th Annual Thanks to generous donations, BSCRF Patient recruitment into International Society for Stem Cell scientists are in Phase III of the human the prostate cancer screening Research meeting. The poster titled, testis stem cell project project is ongoing. The “Onset of Period 2 Oscillation Coincides That the adult human testis contains goal of the project is to develop with Differentiation of Mouse Embryonic pluripotent stem cells, in addition to sperm semen screening tests that Stem Cells" was selected from a record stem cells, was a surprising report by two Dr. Robert Eyre reflect overall male health number of abstracts submitted. It reported research teams a few years ago. Thanks to as well as help diagnose and stage prostate the conclusion that the important circadian private donations, Bedford Research cancer. Recent work suggests that the gene, Period 2, is turned on in stem cells, scientists are determining the efficiency presence of viruses and bacteria in semen but begins to oscillate throughout the with which this naturally occurring source indicates a poorly functioning immune colony when the stem cells begin to of pluripotent cells can be isolated and system. Even mild suppression of immune differentiate. Although the importance of expanded into therapeutically useful, function could allow growth of cancers circadian rhythms to organ function is patient-specific stem cells. They have that would otherwise be naturally rejected. growing in recognition, the Foundation's adopted Good Laboratory Practices for the Fertility and Sterility, the journal of the report was one of only two on circadian testis stem cell derivation to shorten the American Society for Reproductive rhythms at the ISSCR. time to FDA approval of derived lines. Medicine, has recently accepted a report by Bedford Research scientists that Phase III is a collaboration with Dr. Martin cytomegalovirus (CMV) in semen Dym, Georgetown University, who has specimens is related to mildly decreased generously provided cryopreserved immune function in HIV infected men. biopsies of the testis tissues their lab used. CMV is common in humans, usually One of the challenges with testis stem cell causes no symptoms, but can be life- derivation is distinguishing pluripotent Dr. Kiessling (left) answers questions during a threatening to patients receiving bone stem cells (about 500 per gram of tissue) poster session at the meeting. marrow and organ transplants. from the sperm stem cells (about 5 million per gram of tissue) that actively divide to produce many millions of sperm daily -- New Staff Valia Dinopoulou, a one- the proverbial needle in a haystack. If the Alexis Agnew joins the year fellow, hails from the cell dynamics are similar to bone marrow, team as our SPAR coordinator, laboratory of Dr. Dimitrius the pluripotent stem cell is quiescent until bringing over ten years of Loutradis, Professor and activated, in contrast to the sperm stem cell medical experience in both Chairman of Obstetrics, that is actively renewing. Current private practice and the US Gynecology and experiments in the Bedford lab are taking Air Force. SPAR is Reproductive Biology, University of Athens, advantage of this difference to help isolate instrumental in helping couples living with Greece. Valia will work on the project to the stem cells. HIV disease safely parent. genetically engineer parthenote stem cells.

Board Of Trustees Ethics Advisory Board Alan S. Geismer, Esq. Chairman John D. Lee, MEd, Extension Education Arthur Applbaum, PhD, Chairman Robert D. Truog, MD Sugarman, Rogers, Barshak & Cohen Prof. of Ethics and Public Policy Prof of Medical Ethics and Pediatrics, Alan Mayer, AIA Kennedy School of Government Children's Hospital, Harvard Medical School Jose Cibelli, DVM, PhD Principal Architect, Mayer & Associates Prof. of Animal Biotechnology, Stanley J. Bodner, MD Daniel Wikler, PhD, Susan L. Moss, JD, PhD Michigan State University Infectious Disease Specialist Prof. of Ethics and Public Health, Consultant on Equity & Diversity Harvard School of Public Health Lawrence R. Jones Kenneth A. Burry, MD, Anil Purohit, PhD CEO, Jones Marketing Services Prof of Obstetrics and Gynocology Programs for AIDS Testing and Awareness Oregon Health Sciences University Scientific Committee Robert L. Kaufmann, MD Victoria Staebler, CPA Reproductive Endocrinologist Norman Daniels, PhD, Carol M. Warner, PhD, Chairman Senior Financial Advisor, Merrill Lynch Sean Kealy, Esq. Prof. of Ethics & Public Health, Prof. of Biology, Emeritus, Northeastern Boston University School of Law Margaret S. Wray, BSN, Psychiatric Nurse Harvard School of Public Health University

WWW.BEDFORDRESEARCH.ORG 3 BEDFORD STEM CELL Nonprofit Organization Postage and Fees Paid RESEARCH FOUNDATION Bedford Research Foundation PO Box 1028 Zip Code 01730 Bedford, MA 01730 Permit No. 25

News Inside

1 “Off The Shelf” Therapeutic Stem Cells

2 Key Embryo Differences Discovered

3 Update on Stem Cells from Testis

4 New Staff at the Foundation

5 Update on prostate cancer project

BEDFORD STEM CELL RESEARCH FOUNDATION

(Continued from cover) Without the receptor, cells are resistant to positioned to move faster than some HIV infection. traditional academic laboratories because ACTIVATED EGG we are not dependent upon federal funds. SYMPOSIUM Now the task at hand is to create stem cells The parthenote research, highly promising missing CCR5 that are tissue matched to for modifying stem cell genes, cannot be the HIV infected person. Deleting CCR5 in funded by the NIH because of long-time patient-specific (e.g. nuclear transplant, federal funding restrictions. Much of our induced pluripotency, testis or parthenote) laboratory overhead is funded through fee stem cells might work. "Off-the-shelf", for service laboratory testing, allowing Mario Capecchi, PhD engineered stem cells might also work if research donations to go directly to they tissue match the patient. Our 2013 Nov 8, 2013: Dr. Mario Capecchi, Nobel research. We are accountable to individual Activated Egg Symposium brings together Laureate and Distinguished Professor of donors to return the maximum value for pioneers in genetic engineering (Mario every dollar given to the research. Private Human Genetics & Biology at the Cappeccchi and Rudolf Jaenisch) with stem donations -- of all sizes -- are essential to University of Utah School of Medicine, will cell specialists (Treena Arinzeh, Gordon achieving the research speed needed by keynote the tenth Carmichael, Kim Tremblay, Jose Cibelli, patients. annual AES. David DiGiusto) and a member of the Oregon SCNT team, David Battaglia, to Thank you for your Dr. Rudolf Jaenisch, present their work and perspectives in support. Professor of Biology, research areas important to move the work MIT, will present the With gratitude, forward as fast as safely possible to patient dinner “Science Ann A Kiessling, PhD Rudolf Jaenisch therapies. Bedford Research scientists are Perspectives” talk.

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