Developing Targeted Stem Cell Therapeutics for Cancer Shawn Hingtgen, Ph.D. Assistant Professor UNC Eshelman School of Pharmacy May 22 nd, 2013
The Challenge of Drug Delivery for Brain Cancer
Stem Cells in Regenerative Medicine
Stem Cells: Novel Therapeutic Vehicles Stem cells have a unique ability to home to sites of disease in the brain (stroke, injury, cancer) Occurs both after systemic injection and local injection Migration is due to chemotactic signals detected by stem cells Attraction makes them an inherently smart vehicle If armed with drugs, could represent a powerful new therapeutic option Differentiation potential suggests they can regenerate damaged tissue and provide additional curative benefits Tumor death
Targeting Highly Invasive Brain Cancer with Engineered Stem Cells GBM cells=red Stem cells-green
SC-therapy Control Luciferase-TRAIL photon emission (Fold Day 0) Sustained drug delivery: Advantages of Stem Cell-based Drug Delivery 6 5 4 3 IV Infusion Stem Cells Death receptors TRAIL-secreting Stem Cell Tumor death 2 1 0 0 0.5 1 3 7 14 Days Therapeutic Efficacy: Day 1 Day 13 Control Luc-TRAIL
The Challenge of Identifying the Ideal Stem Cell Type for Therapy Immune compatibility with cell-based drugs is major concern Most applications demand a patient s own cells be used (i.e.: personalized medicine) Cells extracted from a patient, armed with therapy, and re-injected Done routinely for stem cell transplants after bone marrow ablation Developing patient-specific neural stem cells for neurologic therapy remains one of the greatest challenges to cell therapy for the brain Stem Cell Source?
Cellular Reprogramming: The New Age of Stem Cell Research Skin biopsy Fibroblast ips Reprogramming genes Neural S.C. Embryonic stem cell Cardiac S.C. Blood S.C. Muscle S.C. Human embryo
Approach: Personalized NSC Therapy for GBM Cellular reprograming offers the potential to realize personalized medicine Biopsy a patient s skin Convert to stem cells Arm with therapies Re-implant therapeutic stem cells for treatment Novel approach to creating personalized NSC-based therapy for GBM Technology to generate reprogrammed NSCs safe for transplant reported only last year These cells have never been used as drug carriers Virtually no reports extending these findings to human reprogrammed NSCs 4. Therapeutic cells reimplanted into patient to cure disease 1. Cells will be removed from a patient s skin 3. Armed with therapies or corrected genetic defect 2. Re-program into desired stem cell type
Mouse Neural Stem Cell Generated Through Cellular Reprogramming Technology Astrocytes Neurons Reprogramming Fibroblast Neural Stem Cell Culture mouse fibroblasts Use cellular reprogramming to convert cells into neural stem cells: inscs Expand in neural stem cell media Cells express neural stem cell markers Differentiate into neural lineage cells Nestin (NSC marker)
Arming Mouse Neural Stem Cell Generated Through Cellular Reprogramming Technology Cell viability (fold control D1) Mo-iNSC-TRAIL inscs modified with lentiviral vectors encoding secreted anti-cancer therapy TRAIL: insc-tr insc-tr robustly express Nestin insc-tr differentiate into astrocytes and neurons insc-tr proliferate at same rate as unmodified insc Astrocytes Neurons 3 2 Control GFPFLuc 1 0 Day 1 Day 3 Day 5 Day 7 Day 10
GBM Cell Viability (Fold Control) Luciferase-TRAIL Signal (Fold Day 1) Mouse insc-tr Induce Killing of GBM Cells In Vitro 1.2 1.0 0.8 0.6 0.4 0.2 0.0 1.2 1.0 0.8 0.6 1 5 10 15 17 Days GFP TRAIL Molecular imaging showed sustained release of anti-cancer therapy by modified cells Modified neural stem cells were cocultured with human GBM cancer cells Tumor cell viability assayed 24 hrs post-stem cell addition Marked decrease in tumor cell viability was observed 0.4 0.2 GBM8 U87 0.0 Control TRAIL
insc-trail Percentage Survival insc-control Photons/sec/cm2 (Fold Day 1) Reprogrammed Mouse Neural Stem Cell Therapies Inhibit Brain Cancer Progression 500.00 400.00 300.00 200.00 100.00 0.00 2x insc-control insc-tr insc-control insc-tr Day 1 Day 3 Day7 Day 14 Day 21 Day 28 10x Reprogrammed mouse neural stem were modified with virus encoding anti-cancer TRAIL or control GFP Mice were implanted with U87 human brain tumors expressing mcherry-luciferase Tumors treated with insccontrol or insc-trail (TR) Serial Fluc imaging was used to track tumor progression 120 100 80 60 40 insc-control insc-trail 20 0 1 8 15 22 29 36 Days
Novel Approach to Generation of Human Neural Stem Cells Human Fibroblast Reprogramming Human Neural Stem Cell Extending research to human cells Developing novel way to generate human neural stem cells using reprogramming technology Very few reports demonstrating this technology Cells express neural stem cell markers Nestin (NSC marker)
Stem Cell-based Therapies: An Eye Towards the Clinics Exploring scaffold systems in mouse models of brain tumor resection Creating novel therapeutics targeting different cancer-specific pathways Immune system regulators Cytotoxic proteins Theranostic multifunctional fusion variants
Acknowledgements Hingtgen Lab Adolfo Alfonso, Ph.D. Julio Rodriguez-Bago, Ph.D. Onyi Okolie, B.S. UNC TraCS Institute -Gene Orringer, M.D. -Susan Pusek Animal Studies Core -Charlene Santos Tissue Culture Facility -Steve Ogelsbee Collaborators Raluca Dumitru, Ph.D. Human Pluripotent Stem Cell Core UNC School of Medicine http://www.med.unc.edu/humanstemcellcore Kam Leong, Ph.D. Department of Biomedical Engineering Duke University Matthew Ewend, M.D. Chair, Department of Neurosurgery UNC Hospitals KL2 Scholars program NC TraCS Pilot Program
Hingtgen Lab Lab: 2070E, Genetic Medicine Office: 2079, Genetic Medicine Phone: 919-537-3827 Email: hingtgen@email.unc.edu