1. "Humanized Mouse model in biomedical science”
Paresh Vishwasrao

2. HSCs Transplantation : Chimeric Reconstitution

3. 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.

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

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

6. Hematopoietic Stem cells Sources for Hu-Mice

7. 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

8.
The Jackson Laboratory

9. 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)

10. Mouse Models and Companies

11. Creating Humanized Mice : Timeline
Humanized Mouse Model
Creating Humanized Mice : Timeline

12. Creating Humanized Mice : Timeline

13. NSG/NRG transplanted with CB stem cells or HSCs

14. Major Mouse Strains from different Labs

15. Human Immune System Models

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

17. 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)

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

19. Peripheral T cell Development

20. The NSG™ Mouse

21. T cell development at 15 wks (Females vs Male)

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

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

24. Limitations of huNSG/huNRG Mice

25. Cytokine Expressed transgenically in NSG mouse

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

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

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

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

30. 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

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

32. The Tumor Xenograft Project : AntiCancer PDOX Pre-Clinical

33. Patient Derived Xenograft (PDX) : mouse model

34. Application of Hu-Mice: Cancer Biologys

35. Summary

36. 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

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

38. Development pathway of Hu-Mice

39. Innate and Adaptive Immune System