Day 2 :
- Cancer Gene Therapy, Advanced Gene Therapeutics
Location: Lone Star West
Juliann G Kiang
Uniformed Services University of The Health Sciences, USA
Texas A&M University Health Science Center
Thomas Bartosh completed his PhD degree in Cell Biology and Genetics from The University of North Texas HSC. He joined the Institute for Regenerative Medicine (IRM) at Texas A&M University in 2008 to develop therapies with mesenchymal stem cells (MSCs). Currently, he an Assistant Professor of Internal Medicine and Director of flow cytometry and microscopy at the IRM. He studies the advantages of using three-dimensional (3-D) culture methods to activate MSCs and exploit their inherent therapeutic potential. This approach was pioneered by Dr. Bartosh at the IRM and has been highlighted in numerous publications.
One appealing approach to cancer therapy involves targeted delivery of exogenous genes encoding an enzyme that converts a specific inert prodrug into cytotoxic derivatives. This strategy, often referred to as targeted suicide gene therapy, has recently been improved by exploiting the high transduction potential and tumor-tracking properties of mesenchymal stem/stromal cells or MSCs. However, clinical applications of tissue-derived MSCs are hindered by donor cell variability and inability to obtain the excessive cell numbers necessary for patient therapies. To overcome these limitations, here we employed induced pluripotent stem cells (iPSCs) as a feeder stock to generate an expandable and uniform source of MSCs. The standardized iPSC-MSCs were engineered to express the suicide gene that codes for cytosine deaminase (CD), an enzyme that converts the non-toxic prodrug 5-fluorocytosine (5-FC) locally into the chemotherapeutic agent 5-fluorouracil (5-FU). These genetically modified iPSC-MSCs constitutively expressed high levels of CD through numerous passages and following cryopreservation, and maintained features characteristic of tissue-derived MSCs. Moreover, the cells showed remarkable ability to kill all cancer cell lines tested (breast, prostate, and ovarian carcinoma, and melanoma) in vitro through bystander effects after addition of 5-FC to the cultures. Level of killing was dependent on time, concentration of 5-FC, and numbers of MSCs used. In mice, both local and systemic injections of iPSC-MSCs expressing CD not only limited formation of human breast tumors following administration of 5-FC, but also triggered regression of established tumors and reduced development of metastatic disease. Interestingly, the anti-tumor effects of iPSC-MSCs were more profound when the cells were primed in culture by addition of 5-FC prior to cell delivery, or suspended in 3-D cultures under conditions that reduced their physical size while increasing uptake by the cancer cells. Importantly, activation of the prodrug also resulted in elimination of the modified iPSC-MSCs thus providing a safeguard against wayward stem cell progeny. Taken together, the results here provide evidence that iPSC-MSCs have immense potential as cellular carriers of therapeutic transgenes and in precision medicine.
Johns Hopkins University School of Medicine, USA
Hiroshi Miyamoto, MD, PhD, is an American board-certified pathologist and is currently an Associate Professor of Pathology and Urology at Johns Hopkins University School of Medicine. He completed his medical school and urology residency training, followed by clinical urology practice, all in Japan. In 1996, he moved to the United States to conduct postdoctoral research. He then completed pathology residency training at University of Rochester Medical Center and clinical fellowship in genitourinary pathology at the Johns Hopkins Hospital. Since 2009, he, as an independent investigator, has been conducting research projects involving molecular biology of steroid hormone receptors in genitourinary tumors
A seminal plasma protein, semenogelin I (SgI), contributes to sperm clotting, upon binding to Zn2+, and can be proteolyzed by prostate-specific antigen (PSA), resulting in release of the trapped spermatozoa after ejaculation. In contrast, the role of SgI in the development and progression of any types of malignancies remains largely unknown. Immunohistochemistry in radical prostatectomy specimens revealed overexpression of SgI in prostatic carcinoma, which was significantly correlated with biochemical recurrence after the surgery. In prostate cancer LNCaP cells, zinc treatment was found to enhance SgI expression. Using prostate cancer cell lines stably expressing SgI, we further investigated its biological functions, in conjunction with zinc (that could generally inhibit cell growth) and androgen/androgen receptor (AR) (that could generally promote cell growth). SgI overexpression resulted in significant increases in AR-positive cell proliferation and PSA expression. Luciferase reporter gene assay showed even slight inhibitory effects of SgI in the absence of zinc versus its significant stimulatory effects in the presence of high levels of zinc on dihydrotestosterone-enhanced AR transactivation. Co-immunoprecipitation then demonstrated dihydrotestosterone-induced physical interactions between AR and SgI. These results suggest that SgI, together with zinc, functions as an AR coactivator and thereby promotes androgen-mediated prostate cancer progression. Additionally, as seen in some of other steroid hormone receptor co-regulators, the LxxLL motif (L=leucine; x=any amino acids) present in SgI appeared to be essential and sufficient for mediating the interaction with AR. Our findings may thus provide a new therapeutic target for androgen-sensitive prostate canceras well as castration-resistant tumor
Iranian Breast Cancer Research Center, Iran
Reza Mehdizadeh received his MSc (2011) in Cell and Molecular Biology from University of Tehran, Iran. He is a researcher at Iranian Breast Cancer Research Center and Ronak Pharmaceutical Company. He is workingon many projects in coordination with cancer National Institute of Genetic Engineering and Biotechnology about molecular targeted therapy especially on triple negative breast cancer. He studied receptor tyrosine kinases (RTKs) and followed their signaling pathways in tumors
The endocannabinoid system consist of cannabinoid CB1 and CB2 receptors, endogenous ligands and the enzymes and processes responsible for the biosynthesis, cellular uptakeor metabolism of these “endocannabinoid” ligands. A wide range of studies were designed to reveal how cannabinoids affects our cell environment: from bone to cancer cells. Endocannabinoid system (ES) as a regulator of bone remodeling, presents novel therapeutic strategies for prevention and treatment of bone disorders. For instance, studies revealed that cannabinoid treatment reduces breast cancer-induced bone loss, fracture, and pain. ES provides an ideal source for cell transplantation or tissue engineering therapies through the positive impact on the survival of differentiated mesenchymal stromal cells (MSCs). Furthermore, Cannabinoid signalingis connected with inhibition of cancer cell proliferation, migration, tumor angiogenesis, and induction of apoptosis through inhibition of p27/KIP1. In contrast, promotion of anti-apoptotic effects and regulation of anti-tumor immune response may induce tumor growth. Some studies indicated that the system’s agonists are effective to regulate breast cancer proliferation, invasion, and migration. On the other side, some evidences showed that in the absence or lowconcentrated cannabinoid receptors in tissues, the administration of synthetic and/or phytocannabinoids may increases incident of cancers. We need to determine the balance between these diverse mechanisms, and how they modulate cancer in vivo. In conclusion, the endocannabinoid system is an exciting target for research on the treatment of cancers, especially triple negative breast cancer which has poor prognosis without any standard targeted therapy. By further clinical trials and modern genome engineering methods, cannabinoid system has a great potential for studying therapeutic options in the future
Harvard Medical School, USA
Daniela Dinulescu is an Assistant Professor at Harvard Medical School. She received her PhD from Oregon Health and Science University and completed her Postdoctoral studies in the field of Cancer Genetics at MIT. Her research interests focus on cancer biology, malignancies of the gonads and reproductive tract, with a special emphasis on ovarian cancer research and endometriosis. Our laboratory is actively investigating the key contribution of cancer stem cells (CSCs) to tumor chemoresistance. Our current studies focus on better understanding the mechanism of stem cell signaling in the maintenance of the CSC niche and ovarian tumorigenesis. The aim is to harness the power of nanotechnology to develop improved “homing” technologies for the delivery of therapeutic agents specifically targeting and sensitizing ovarian cancer cells, including CSCs, in a spatio-temporal fashion
Ovarian cancer is the fifth leading cause of cancer death in women. An exciting hypothesis emerging in cancer biology is that cancer cells with stem cell properties are a major driving force for both tumor development and chemoresistance to therapy based on their propensity to divide indefinitely and survive standard cancer chemotherapy. This could explain the high rate of therapeutic failure and tumor relapse seen in patients. Our research group has recently generated animal models of ovarian cancer that recapitulate the human disease with great accuracy. Our animal models constitute invaluable tools that will help identify, characterize, and define the key roles of ovarian cancer stem cells in tumor metastasis and resistance to chemotherapy. Furthermore, they provide us with unique, relevant systems in which to screen new and exciting therapies specifically targeting ovarian cancer stem cells. Our current investigation focuses on whether ovarian cancer stem cells contribute to platinum chemoresistance and tumor relapse. In addition, we are examining whether key stem cell markers constitute good markers for predicting chemoresistance to platinum and doxorubicin in ovarian cancer. Finally, we plan to test a nanotechnology-based strategy for therapeutic delivery, which will selectively target and sensitize cancer stem cells in a spatiotemporal fashion. This constitutes a novel paradigm in the strategies towards developing effective therapies and a cure for ovarian cancer. It is well established that the chemotherapeutic options used today are of limited value in ovarian cancer patients who relapse. The addition of a front-line therapeutic agent specifically targeting cancer stem cells have the potential to greatly improve the high rate of therapeutic failure and tumor relapse currently seen in ovarian cancer patients and especially platinum resistant or refractory patients worldwide. Our studies aim to reduce cancer mortality rates and improve the quality of life of cancer patients through more efficient and less toxic therapies. Data collected from these key preclinical studies will be instrumental in the design of new human clinical trials for ovarian cancer therapy selectively targeting cancer stem cells
Kuwait Medical Genetic Center, State of Kuwait
Laila Ali Bastaki completed PhD and currently working as a Director of Kuwait Medical Genetic Center at State of Kuwait.
Duchenne muscular dystrophy (DMD) is a severe muscle wasting disorder, affecting around 1 in 3500 newborn boys, caused by mutations in the Dystrophin gene. The disease is characterized by complete loss of muscle dystrophin protein causing progressive muscle weakness leading to premature death due to heart and respiratory failure. Two gene therapy modalities, Antisense oligonucleotides (AONs) for frameshift deletions and ataluren for nonsense mutations, are recently introduced to treat the condition. The rationale behind these therapeutic modalities is to restore the reading frame of the Dystrophin gene and produce a partially functional dystrophin protein isoforms in skeletal muscle. In Kuwait we registered about 100 DMD patients in our institute neuromuscular registry. All patients were confirmed by MLPA or full gene sequencing. We revised the registry and found seven of our patients legible for AONs and three for ataluren therapy. These patients will be included soon in the treatment program supported by Kuwaiti government. In this presentation, I will discuss our experience regarding DMD natural history and the gathered experiences about gene therapy of the selected cases.
Kiev regional p/n hospital, Ukraine
Ponizovskiy M R has graduated from N.I.Pirogov Vinitsa National Medical University. He worked manager of the hospital in Sum district of Ukraine and then as a Doctor of therapeutic department in the Kiev regional hospital. Then he has graduated from Kiev national Institute of post diploma education and Lvov National Medical University, faculties of laboratory clinical diagnostics, laboratory clinical biochemistry, laboratory toxicology. In 1972 – 1990, he worked as the head of clinical and biochemical laboratory of the First Kiev regional hospital, and I have obtained Degree PhD from 1990. In 1990 – 2002, he worked as the head of toxicological and biochemical Laboratory of the Kiev regional p/n hospital. He has the scientific degree PhD and the clinical degree of the highest category doctor. In Germany, he is the member of the Society of Inventors: he has the German protection of the invention [Gebrauchsmusters] of the new method of blood cells separation and allocation, and this method is examined in German Patent Department for reception of the German Patent.
Balance catabolic exoergonic and anabolic endoergonic processes, causing chemical potentials and electric charges on cellular membranes exerting cellular capacitors operation, is the mechanism maintenance stability Internal Energy [36.6°C by which all enzymes operate etc.] of an organism, according first law of thermodynamics. Shift balance catabolic & anabolic processes into excess catabolic processes leads to inflammatory and infection diseases. Shift balance catabolic and anabolic processes into excess anabolic processes leads to proliferative processes of Cancer, Leukemia etc. Having borrowed from the folk healers Omelchenko A. and Breusse R. the method treatment of the oncologic patients, having tested the positive results of this method of cancer treatment, the mechanism of this method treatment was explained and was substantiated this method treatment, using the offered concepts of Warburg effect mechanism and mechanism transmutation in mitochondria function in cancer genesis. The author was convinced of the efficiency of this method cancer treatment by the meetings with cured patients and determined on own experience the efficiency of treatment the ill man with the incurable cancer stage. The mechanism of this method of cancer therapy operates via Warburg effect targeting which leads to cancer metabolism depression. The new method Cancer Therapy proposes combination “Prolonged medical starvation” with considerably decreased dosage of cytotoxic drugs. The detailed scientific explanation of mechanism operation cancer treatment method via “Prolonged medical Starvation 45 days” was based on the offered concepts of Warburg effect mechanism which exhibits cancer metabolism. Here is the short simple explanation of this method treatment mechanism: The forced inflow of substances and energy in condition of prolonged medical starvation occurs from the organism’s depots both for the organism metabolism and for the cancer metabolism. The exhausted organism’s depots in condition of prolonged medical starvation exerts shift of balance catabolic and anabolic processes of the organism into catabolic processes for maintenance stabile Internal Energy of an organism (temperature 36.5ºC – 37.2ºC, by which all enzymes operate) that causes shift also tumor metabolism into catabolic pathway violating anabolic processes in tumor metabolism. Thus inhibition of the abundance anabolic processes, characterized tumor metabolism, results in tumor depression. Thereby the treatment with considerably decreased dosage of cytotoxic drugs damage tumor metabolism and rearrange Warburg effect into Pasteur effect, causing cancer depression. Just depressed cancer tumor is efficiently destroyed by the decreased dosage cytotoxic drugs. The advantage of the new method of cancer treatment is that the new method of cancer disease treatment does not intrude into the stability of Internal Medium and Internal Energy an organism and cells of an organism, does not violate defensive mechanisms of an organism /immune and hormonal systems/in comparison with targeting metabolic links of anabolic processes both in the organism and in the tumor, causing damage of hormonal regulatory processes and protective immune processes in an organism by up-to-date chemotherapeutic methods which use great dosage cytotoxic drugs. Therefore the efficient method cancer therapy using decreased dosage of cytotoxic drugs against depressed cancer tumor does not lead to negative consequences how recurrence cancer disease after some medical remissions, resistance to cytotoxic drugs after long anticancer therapy etc. Also it was suggested the possible modes to integrate the offered method treatment of cancer disease with the modern methods treatment of cancer disease which should be made after detailed clinical trials
Arizona State University, USA
Samira Kiani is an Assistant Professor in the school of biological and health systems engineering at Arizona State Univeristy. She completed her Postdoctoral training in the center for Synthetic Biology at Massachusetts Institute of Technology, where she worked on developing synthetic gene circuits to reprogram the function and behavior of mammalian cells based on the Clustered Regularly Interspaced short Palindromic Repeats (CRISPR)/Cas9 technology. Her lab is interested in developing synthetic gene circuits using the Cas9/gRNA engineering strategies to edit/modulate endogenous genes or transgenes in human cells towards reprogramming cellular fate/function.
Cas9 is an RNA guided DNA endonuclease of the bacterial Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR) system that has been adopted for programmable genome editing and transcriptional regulation. As the complexity of genetic interrogations with this technology is rapidly growing, there will soon be a need for safer and more specific CRISPR-based therapies. To this end, we apply the design principles of synthetic biology to develop genetic logic gates, layered genetic circuits and kill switches, which allow us to exert internal control over the function of the CRISPR system. We develop RNA Pol II driven gRNAs that enable cell and context specific expression of them. This strategy combined with synthetic promoter engineering allow us to develop genetic circuits to have more specific CRISPRs and avoid off target effects. In parallel, we have devised a strategy to readily switch between catalytically active and null Cas9 protein, allowing knocking out and activating target genes in the same cells using distinct gRNAs. By truncating the gRNA from 5’ end, we have shown that gRNAs with 14nt guide sequences ablate the nuclease activity of Cas9, while leaving its DNA targeting capacity intact. By fusing a Cas9 nuclease protein to a potent activation domain, VPR or application of gRNAs with MS2 binding loops, we can use a same Cas9 protein for transcriptional activation (14nt gRNAs) or gene disruption (20nt gRNAs). By incorporating this capacity within our layered circuits, we have developed and tested several synthetic gene kill switches that can modulate the transcriptional activity of Cas9 protein. Considering the growing clinical application of DNA-based gene therapies and vaccines, the developed CRISPR logic circuits pave the way towards developing safer and more specific gene therapies and allow us to perform more sophisticated biological discoveries
Shahrekord University of Medical Sciences, Iran
Mohammad-Saeid Jami has completed his PhD from the University of Leon and postdoctoral studies from University of Nebraska Medical Center (NE) and St. Johns University (NY). He is assistant Professor at the department of cell therapy at Shahrekord University of Medical Sciences (SKUMS). He has published more than 12 papers in ISI journals and has been serving as an editorial board member of Journal of Cancer and Clinical Oncology and Journal of Molecular Biology Research
Hearing loss (HL) is one of chronic diseases with high prevalence. In addition to congenital and age-related hearing loss, sensorineural hearing loss occurs after mechanosensory hair cells (HC) injury due to ototoxic agents, trauma and noise. In the mammalian cochlea, the organ of Corti contains of HCs wherein Atoh1 is one of the most important determinants whose function is required in early stages of development of inner ear HCs. miRNA-183 family including miR-96, miR-182 and miR is highly expressed in hair cells and is known to be directly involved in HC differentiation. Here we compared overexpression of Atoh1 and miR-183 family as two different strategies to differentiate HCs from both human Mesenchymal Stem Cells (hMSCs) and Hair Follicle Pluripotent Stem Cells (hfPSCs) as two cellular sources. The hfPSCs seem to be a better candidate for differentiation of HCs and expression of hair cell protein markers and it may be due to the ectodermal origin of this cells