Stem Cell Therapies 2024 PDF

KIMN10 Dr. Agathees Subramaniam Associate Professor Div. of Molecular Medicine and Gene Therapy November 2024 STEM CELL THERAPIES: CURRENT APPLICATIONS AND CHALLENGES LEARNING OUTCOMES ¡ Define what stem cells are ¡ Classify the...

KIMN10 Dr. Agathees Subramaniam Associate Professor Div. of Molecular Medicine and Gene Therapy November 2024 STEM CELL THERAPIES: CURRENT APPLICATIONS AND CHALLENGES LEARNING OUTCOMES ¡ Define what stem cells are ¡ Classify the different stem cell types ¡ Describe how stem cells can be used therapeutically ¡ Discuss current limitations for stem cell therapies SCAN ME (LINK TO PADLET) LECTURE OVERVIEW ¡ PART 1 ¡ What are stem cells? ¡ Stem cell classification ¡ Pluripotent stem cells: ES cells and iPS cells ¡ Multipotent stem cells ¡ PART 2 ¡ Translation to the clinic: several considerations ¡ Current applications of Stem Cell Therapies  STEM CELL THERAPY: REGENERATIVE MEDICINE š Why? š Shortage of donated organs š Incompatibility of available donations š How? š Repairing tissues/organs rather than treating symptoms Data from US. š Replacing defective/diseased cells with new functional cells š Which sources? š Isolated or induced stem cells European Directorate for the Quality of Medicines & HealthCare https://www.edqm.eu/en/eodd  WHAT ARE STEM CELLS? Stem cells have two defining qualities: š They can self-renew š They can differentiate into specialized mature cells Modified from Doulatov et al., 2012, Cell Stem Cell  WHAT IS THE ORIGIN OF STEM CELLS? Zakrzewski et al. Stem Cell Research & Therapy (2019); Pohl, H. www.enzolifesciences.com  STEM CELL CLASSIFICATION ¡ Totipotent stem cells can give rise to an organism: e.g. a zygote ¡ Pluripotent stem cells can give rise to all three germ layers (mesoderm, endoderm and ectoderm) ¡ Multipotent stem cells = Valuable for research and clinical applications. adult stem cells, e.g. a neural stem cell or a hematopoietic stem cell ISSCR, Stem cell facts  EMBRYONIC STEM CELLS (ES CELLS) š Murine ES cell lines were first established in 1981. š Human ES cell lines were first derived in 1998. š ES cells can be isolated from excess in vitro fertilized (IVF) eggs which have been donated for research purposes. Evans, M.J., and Kaufman, M.H. (1981). Establishment in culture of pluripotential cells from mouse embryos. Nature 292, 154– 156. Thomson, J.A., Itskovitz-Eldor, J., Shapiro, S.S.,Waknitz, M.A., Swiergiel, J.J., Marshall,V.S., and Jones, J.M. (1998). Embryonic stem cell lines derived from human blastocysts. Science 282, 1145–1147. Hasegawa 2010  ES CELLS AND THEIR POTENTIAL š ES cells can proliferate indefinitely and maintain their pluripotent state. š ES cells can give rise to all three germ layers. To this day, ES cells have been successfully converted to neurons, blood cells, cardiomyocytes and liver cells, among others. š Crucial for the in vitro study of developmental processes which cannot be assessed otherwise š Disease modelling (genetic diseases, human host-pathogen interactions, organoid research, etc…) š Great potential for advancement of cell therapy = transplantation of cells of interest differentiated from ES cells into patients  LIMITATIONS OF ES CELLS š Ethical concerns about the human blastocysts which have to be sacrificed to obtain ES cells. This led to a prohibition of work with human ES cells in some countries or, where allowed, this work is tightly regulated š As donor ES cells derive from a different organism, allogeneic transplantation to a patient could lead to immune rejection due to unmatched human leukocyte antigen (HLA) haplotypes.  LIMITATIONS OF ES CELLS š Steps to circumvent immune rejection: š Biobanking: creation of an ES cell bank with ES cell lines derived from HLA haplotypes commonly found in the population š 150 selected homozygous HLA-typed cell lines could match 93% of the UK population while 140 unique HLA homozygous donors would be needed to cover 90% of the Japanese population. š Somatic cell nuclear transfer (SCNT): production of ES cells from the patients themselves by therapeutic cloning SOMATIC CELL NUCLEAR TRANSFER (SCNT) Gurdon 2009 SOMATIC CELL NUCLEAR TRANSFER (SCNT) John B. Gurdon receiving the Nobel Prize in Physiology or Medicine in 2012 “for the discovery that mature cells can be reprogrammed to become pluripotent”  SOMATIC CELL NUCLEAR TRANSFER (SCNT) Dolly  THERAPEUTIC CLONING Colman and Kind (2000) Trends in Biotechnology  INDUCED PLURIPOTENT STEM CELLS (IPS CELLS): A TURNING POINT š In 2006, Shinya Yamanaka’s lab show that fully differentiated somatic cells can be “reprogrammed” to a pluripotent stem cell state. Takahashi, K., Tanabe, K., Ohnuki, M., Narita, M., Ichisaka, T., Tomoda, K., and Yamanaka, S. (2007). Induction of Pluripotent Stem Cells from Adult Human Fibroblasts by Defined Factors. Cell 131, 861–872. Takahashi, K., and Yamanaka, S. (2006). Induction of Pluripotent Stem Cells from Mouse Embryonic and Adult Fibroblast Cultures by Defined Factors. Cell 126, 663– 676. ISSCR, Stem cell facts  INDUCED PLURIPOTENT STEM CELLS (IPS CELLS): A TURNING POINT š In 2006, Shinya Yamanaka and colleagues show that fully differentiated somatic cells can be “reprogrammed” to a pluripotent stem cell state. Takahashi, K., Tanabe, K., Ohnuki, M., Narita, M., Ichisaka, T., Tomoda, K., and Yamanaka, S. (2007). Induction of Pluripotent Stem Cells from Adult Human Fibroblasts by Defined Factors. Cell 131, 861–872. Takahashi, K., and Yamanaka, S. (2006). Induction of Pluripotent Stem Cells from Mouse Embryonic and Adult Fibroblast Cultures by Defined Factors. Cell 126, 663– 676. ISSCR, Stem cell facts DISCOVERY OF IPS CELLS https://www.pbslearningmedia.org/resource/nsn08.sci.life.stru.stemcell2/stem-cells-breakthrough/ IPS CELLS AND THEIR POTENTIAL ¡ All the advantages of ES cells, and also: ¡ No need for embryos/No ethical issues ¡ Vastly renewable. Cell number is not a limitation. iPSC cell lines can be made and stored ¡ Easily accessible. Variety of somatic cells such as blood, keratinocytes, fibroblast and hepatocytes can be used ¡ Individual-specific i.e. personalized or non-immunogenic IPS CELLS AND THEIR POTENTIAL Volarevic et al. - 2018 LIMITATIONS OF IPS CELLS ¡ Risk of transplanting incompletely differentiated cells ¡ Genomic and epigenetic alterations in the iPSC-derived cells ¡ Tumorigenicity due to the incomplete silencing of the oncogenic transcription factors (Myc and Klf4) ¡ Personalized medicine is not practically feasible ¡ In 2014, the autologous transplantation of iPSC-derived retinal pigment epithelium (RPE) cells required the patient to wait over 10 months for the surgery and cost nearly US $1 million [Umekage et al. – 2019] MULTIPOTENT STEM CELLS ISSCR, Stem cell facts  MULTIPOTENT STEM CELLS ¡ They are tissue- specific ¡ Hard to access and isolate  HEMATOPOIETIC STEM CELLS (HSCS) ¡ Most widely transplanted cells in stem cell therapies ¡ Transplanted HSCs home to the bone marrow ¡ HSCs replenish the recipient’s hematopoietic system Manz & Boettcher Nat Rev Immunology (2014) HISTORY OF HSC TRANSPLANTATION ¡ Aftermath of WW II: Ionizing radiation induced death was reversed by transplanting bone marrow from healthy mice. ¡ 1956: First successful stem cell transplant by Dr. E. Donnall Thomas to treat leukemia in a patient who had a perfect donor from the identical twin CONSIDERATIONS FOR HSC TRANSPLANTATION ¡ 1970s: Graft versus host disease (GVHD) ¡ 1980s and 1990s: Conditioning regimens ¡ Sources: ALLOGENEIC VS AUTOLOGOUS HSC TRANSPLANTATION INDICATIONS FOR HSC TRANSPLANTATION Henig and Zuckerman - 2014  ALLOGENEIC HSC TRANSPLANTATION ¡ Donor T cells and conditioning regimen eliminate leukemia cells ¡ Patient transplanted with a new hematopoietic system ¡ Graft versus leukemia (GvL) effect  MESENCHYMAL STEM CELLS (MSCS) ¡ MSCs can differentiate in vitro into osteocytes, chondrocytes and adipocytes ¡ The markers they express are still widely debated ¡ Almost 1,000 registered clinical trials with the use of MSCs on clinicaltrials.gov Volarevic et al. - 2018  MESENCHYMAL STEM CELLS (MSCS) currently in use in the clinic ¡ Anti-inflammatory properties, influence on tissue repair ¡ Short-term existence: allogeneic safety ¡ Current MSC clinical trials utilize their immune suppression properties ¡ Keep in mind: significant diversity in how MSCs are described and manufactured Andrzejewska et al. - 2019 LECTURE OVERVIEW ¡ PART 1 ¡ What are stem cells? ¡ Stem cell classification ¡ Pluripotent stem cells: ES cells and iPS cells ¡ Multipotent stem cells ¡ PART 2 ¡ Translation to the clinic: several considerations ¡ Current applications of Stem Cell Therapies TRANSLATION TO THE CLINIC What do we need to consider when taking a stem cell therapy to the clinic? SCAN ME (LINK TO PADLET) TRANSLATION TO THE CLINIC ¡ How to overcome the scarcity of stem cells? ¡ How to design protocols to obtain the cells of interest from stem cells? ¡ How to assess the safety of the material to be transplanted? ¡ How to efficiently deliver the cells? Madl 2018 TRANSLATION TO THE CLINIC ¡ How to overcome the scarcity of stem cells? ¡ How to design protocols to obtain the cells of interest from stem cells? ¡ How to assess the safety of the material to be transplanted? ¡ How to efficiently deliver the cells? Madl 2018  HOW TO OVERCOME THE SCARCITY OF STEM CELLS? ¡ Extracting from alternate sources: ¡ Example from cord blood ¡ Contains mesenchymal and hematopoietic stem cells Cord blood advantages Increased donor availability Non-invasive process Lower risk of GvHD Reduced need for HLA matching Cord blood disadvantages Limited HSC number  HOW TO OVERCOME THE SCARCITY OF STEM CELLS? ¡ Expanding hematopoietic stem cells ex vivo:  HOW TO OVERCOME THE SCARCITY OF STEM CELLS? ¡ Expanding hematopoietic stem cells ex vivo: UCB HSCs Knockdown validation: LSD1 inhibition promotes HSPCs in vitro Western Blot qPCR ditions Weekly FACS analysis HSC expansion Zemaitis et al., 2023, JBC Subramaniam et al., 2020, Blood Patient Subramaniam et al., 2019, Hematologica TRANSLATION TO THE CLINIC ¡ How to overcome the scarcity of stem cells? ¡ How to design protocols to obtain the cells of interest from stem cells? ¡ How to assess the safety of the material to be transplanted? ¡ How to efficiently deliver the cells? Madl 2018  BLOOD FORMATION DURING DEVELOPMENT AGM = Aorta-Gonad-Mesonephros EMPs = Erythro-Myeloid Progenitors Yoder Nat Biotech (2014) BLOOD FORMATION DURING DEVELOPMENT Emerging HSCs Endothelial to hematopoietic Hemogenic transition (EHT) Endothelium (HE)  HOW TO DESIGN PROTOCOLS TO OBTAIN THE CELLS OF INTEREST FROM STEM CELLS? ¡ cGMP-compliant protocol ¡ Ensure complete differentiation of all pluripotent stem cells ¡ Make sure the cells of interest are obtained (assess markers) ¡ Obtained cells tested for functionality TRANSLATION TO THE CLINIC ¡ How to overcome the scarcity of stem cells? ¡ How to design protocols to obtain the cells of interest from stem cells? ¡ How to assess the safety of the material to be transplanted? ¡ How to efficiently deliver the cells? Madl 2018  HOW TO ENSURE THE SAFETY OF THE MATERIAL TO BE TRANSPLANTED? Risk: Teratomas Subcutaneous injection  HOW TO ENSURE THE SAFETY OF THE MATERIAL TO BE TRANSPLANTED? Risk: Insertional Mutagenesis Moradi et al. - 2019  HOW TO ENSURE THE SAFETY OF THE MATERIAL TO BE TRANSPLANTED? Genomic integrity Tapia and Schöler 2016 HOW TO ENSURE THE SAFETY OF THE MATERIAL TO BE TRANSPLANTED? ¡ iPSC quality control check list: ¡ Short tandem repeat analysis ¡ Identity analysis ¡ Residual vector testing ¡ Karyotype ¡ Viral testing ¡ Bacteriology ¡ Single nucleotide polymorphism arrays ¡ Flow cytometry ¡ Phenotypic pluripotency assays ¡ Histone modification and DNA methylation Zakrzewski et al 2019 TRANSLATION TO THE CLINIC ¡ How to overcome the scarcity of stem cells? ¡ How to design protocols to obtain the cells of interest from stem cells? ¡ How to assess the safety of the material to be transplanted? ¡ How to efficiently deliver the cells? Madl 2018  HOW TO EFFICIENTLY DELIVER THE CELLS? Transdifferentiation Graf, 2013 Mollinari 2018  HOW TO EFFICIENTLY DELIVER THE CELLS? Organoids Rossi 2018  HOW TO EFFICIENTLY DELIVER THE CELLS? Bioengineering Hasan et al 2016 PARKINSON’S DISEASE Pioneered in Lund over 30 years ago by A. Björklund & O. Lindvall, using fetal cells obtained from the midbrain of aborted embryos Follow-up 24 years later showed that transplanted neurons engrafted and survived (Li et al, PNAS 2016) Henchcliffe and Parmar - 2018  A NEW SC THERAPY FOR PARKINSON’S DISEASE ¡ STEM-PD: a stem cell-based therapy for the treatment of Parkinson’s Disease from Lund University ¡ Regulatory approval for a Phase I/IIa clinical trial October 2022 ¡ Human ES cell based dopamine nerve cell product M. Parmar and A. Kirkeby ¡ The trial plans to enroll a total of 8 patients for transplantation, starting with patients from Sweden https://www.stemcellcenter.lu.se/article/swedish-medical-products-agency-grants-approval-clinical-study-new-stem-cell-based-parkinsons  IPSC-DERIVED CELL THERAPY TRIALS ¡ Currently there are 19 registered clinical trials involving iPSC-derived cell therapy (interventional) ¡ Various diseases are targeted by these cell therapies Kim et al – 2022 – Stem Cell Rev and Rep IPSC-DERIVED CELL THERAPY PRODUCTS ¡ Companies aiming to develop iPSC-derived cell therapy products ¡ Several have registered clinical trials Kim et al – 2022 – Stem Cell Rev and Rep STEM CELL CLINICS?  RECOMMENDED READING Reviews Hoang, D. M. et al. Stem cell-based therapy for human diseases. Sig Transduct Target Ther 7, 1– 41 (2022). Hu, J., and Wang, J. (2019). From embryonic stem cells to induced pluripotent stem cells— Ready for clinical therapy? Clinical Transplantation 33, e13573. Moradi, S., Mahdizadeh, H., Šarić, T., Kim, J., Harati, J., Shahsavarani, H., Greber, B., and Moore, J.B. (2019). Research and therapy with induced pluripotent stem cells (iPSCs): social, legal, and ethical considerations. Stem Cell Research & Therapy 10, 341. LEARNING OUTCOMES ¡ Define what are stem cells ¡ Classify the different stem cell types ¡ Describe how stem cells can be used therapeutically ¡ Discuss current limitations for stem cell therapies SCAN ME (LINK TO SLIDO) Pluripotent stem cells can give rise to neural stem cells. True or False? ESCs and iPSCs are equivalent for transplantations as part of stem cell therapies. True or False?  Umbilical cord blood derived stem cells can be used for clinical applications directly True or False? Induced pluripotent stem cells pose no concern of immune rejection. True or False? Guided differentiation of induced pluripotent stem cells into blood cells in a lab setting can be used for clinical applications. True or False?