"Humanized Mouse model in biomedical science” Paresh Vishwasrao

HSCs Transplantation : Chimeric Reconstitution

Introduction of Humanized Mouse Model What, how, when and why ? Creating Humanized Mice : Timeline Development and Functional Aspect. Limitations and Improvisation. Applications of Humanized Mouse Model Patient Derived Xenograft (PDX) : mouse model.

Humanized Mouse Model : hu-Mice What, how, when and why ?

“Hu-Mice”: Mice with human HSCs Pre-Clinical Models Cancer Infections Transplantations Pharm/Tox Stem cells, NIH (2001)

Hematopoietic Stem cells Sources for Hu-Mice

Translating lessons from clinics Source and selection of the stem cells (No T cells) Matching of the human histocompatibility complex Depletion of the “original” hematopoietic system Period of immune deficiency (6-12 months) Graft versus host disease

https://www.youtube.com/watch?v=lHo2hEKCvSA The Jackson Laboratory

Factors determining Human Immune Responses Age of the recipient Strain background and transgenic factors Source of stem cells and human tissues Cell dose and percentage of sub populations Route of administration Sub-lethal radiation dose Surgical procedure and safe housing Brehm et al Clin. Immunol (2010) Vishwasrao.P and Sykes M, ASH (2014)

Mouse Models and Companies

Creating Humanized Mice : Timeline Humanized Mouse Model Creating Humanized Mice : Timeline

Creating Humanized Mice : Timeline

NSG/NRG transplanted with CB stem cells or HSCs

Major Mouse Strains from different Labs

Human Immune System Models

Development and Functional Aspect. Humanized Mouse Model : hu-Mice Development and Functional Aspect.

Engraftment of human Immune System Develops robust Immune system of multiple linages. High T cell and low B cell development. T cell education through autologous thymic tissue. Detectative viral infections (EBV and HIV). Rejection of Pig islets. Nichole Wash, L Shultz. Nat Rev Imm (2017)

Thymus development in BLT mice at 16 Wks post-implant Michael Brehm and Shultz. Nat Med (2008)

Peripheral T cell Development

The NSG™ Mouse

T cell development at 15 wks (Females vs Male)

T cell in PBL< Spleen, LN at 20Wks(Females vs Male)

Limitations and Improvisation Humanized Mouse Model : hu-Mice Limitations and Improvisation

Limitations of huNSG/huNRG Mice

Cytokine Expressed transgenically in NSG mouse

Making of Humanized Mouse Model: “Hu-Mice” i.v/i.p/s.c/i.t Vishwasrao.P and Sykes.M. Am J Transplant (2017)

NSG-SGM3: Stem cell factor, GM-CSF and IL3 New Born , Fetal liver CD34+ and intrahepatically Billerbreck, Ploss et al, Blood (2011)

MISTRG: M-CSF, IL3, SIRPa, THPO and GM-CSF New Born , Fetal liver CD34+ and intrahepatically Rongvaux, Manz, Flavellet al, Nat Biot (2014)

Applications of Humanized Mouse Model Humanized Mouse Model : hu-Mice Applications of Humanized Mouse Model

Application of “Hu-Mice” Infectious disease: Typhoid Fever/Salmonella, Mycobacterium, Malaria, Sepsis Regenerative Medicine: Stem cell transplant Immunity and Autoimmunity: Type 1 Diabetes, Allergic inflammation Transplantation: Xenotransplantation Human Hematopoiesis: Immunobiology, GVHD Cancer Biology: PDX, NK cell therapy, T cell editing

Patient Derived Xenograft (PDX) : mouse model. Humanized Mouse Model Patient Derived Xenograft (PDX) : mouse model.

The Tumor Xenograft Project : AntiCancer PDOX Pre-Clinical https://www.youtube.com/watch?v=DWYkC3CEVB8

Patient Derived Xenograft (PDX) : mouse model

Application of Hu-Mice: Cancer Biologys

Summary

Acknowledgement PhD Lab-Sykes lab Prof. Megan Sykes Previous- PhD lab Dr. HaoWei Li Previous- PhD lab Prof. Christian Buske PhD lab Dr. C.W .Choi Postdoc lab- Davila lab Prof. Marco Davila Dr. Gongobo Collaborator Prof. Marcel Maus Prof. David Weinstock

Schematics : of Immunodeficient mouse models Leonard Shultz. Nat Rev Imm (2007)

Development pathway of Hu-Mice

Innate and Adaptive Immune System