Lisbeth A' Welniak, PhD

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Lisbeth A. Welniak, PhD Research Assistant
Professor p40 Cytokines in Allogeneic BMT Graft
versus Host Disease and Anti-Tumor Activity
Networking Scientists and Resources to
Strengthen Biomedical Research in Nevada
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Allogeneic Hematopoietic Stem Cell Transplantation

• Treatment for neoplasia and diseases involving
  hematopoietic cells.
• Leukemias
• Lymphomas
• Solid Tumors (i.e. renal, ovarian)
• Myelodysplastic Disorders
• Non-Malignant Diseases
• Aplastic Anemia
• Variety of Inherited Disorders
• Autoimmune diseases

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Benefits and Problems of Allogeneic Transplants

• Benefits
• Only available curative treatment for a number of
  diseases
• Problems
• Relapse of disease in a percentage of transplants
• Significant morbidity and mortality from
  treatment related complications
• Graft-versus-Host Disease (GVHD)
• Opportunistic Infections
• Graft rejection
• Organ Toxicity

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Pathobiology of GVHD
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Cytokines in T Cell Differentiation and Function

• Interleukins 12 and 23
• Produced by dendritic cells
• Heterodimers consisting of a common (p40) and an
  unique protein subunit
• Drive the development of Th cell subset
  differentiation
• Interferon-g and Interleukin-17
• Effector cytokines that define two T cell subsets

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Emerging Paradigm of Cellular CD4 (helper) T
Cell Subsets
GVHD
Th17 - Autoimmune disease effector
Th0
GVHD GVT
Th1 Classic cell-mediated effector
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Hypothesis

• Host-derived p40 cytokines (IL-12 and IL-23) are
  critical mediators of GVHD.
• Host-derived IL-12 and IL-23 will differentially
  generate Th1 and Th17 cells in GVHD.
• Both Th1 and Th17 cells are cellular mediators of
  Graft versus Host Disease (GVHD). Th1 cells, not
  Th17 cells, are critical for establishment of
  long-term memory Graft-versus-Tumor (GVT)
  responses.

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Specific Aims

• Specific Aim 1
• Compare allo-reactive T cell activity following
  induction of acute GVHD in wild type (WT), IL-12
  p35 KO and p40 KO mice. Will determine
• the extent of donor T cell proliferation and
  alloreactive T cell expansion
• cytokine production (i.e. Th1, Th2, Th17
  cytokines) and CTL activity
• the GVHD-associated histopathology of the
  multiple target organs.
• Specific Aim 2
• Determine if GVH and GVT activities are similarly
  influenced by the IL-12 family of cytokines
  through use of WT, IL-12 p35 KO and p40 KO mice.

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Allogeneic BMT Model for GVHD/GVT
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Experimental Tools

• Mouse Strain IL-12 IL-23
• Wild Type (WT)
• IL-12 p35 KO -
• IL-12 p40 KO - -

Previous studies have shown that neutralization
of p40 reduces GVHD. I have shown this is
primarily due to p40 cytokines produced by the
recipient.
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Accelerated GVHD Lethality in p35 KO Recipients
Conclusion Severity of acute lethal GVHD is
increased in animals in which host APCs can
produce IL-23 but not IL-12.
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Summary of Progress Specific Aim 1

• Data suggests the small intestine may be
  preferentially targeted in GVHD in mice lacking
  IL-12 (IL-23 present).
• Results demonstrate serum IFNg and TNFa
  production is delayed in recipient mice lacking
  IL-12 (IL-23 present).
• IFNg limits alloreactive T cell proliferation in
  GVHD.

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Current Directions

• Expand evidence for Th1 and Th17 subset
  involvement in GVHD (i.e. IL-17 production)
• Develop tools in study IL-23 in non-genetically
  modified mice (i.e. siRNA and gene delivery)
• Determine the role of Th1 cells and Th17 cells in
  Graft-versus-Tumor (GVT) responses.

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INBRE Supported Publication

• Welniak LA et. al., Blood, 107410-2. 2006.
• Peyer patches are not required for acute
  graft-versus-host disease after myeloablative
  conditioning and murine allogeneic bone marrow
  transplantation.
• This study also demonstrated that recipient
  derived TNFa (a cytokine induced by IL-17) is not
  essential for development of GVHD.

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Grant Submissions July 2005 June 2006

• Pending
• R15-AT003591 (Running Welniak, co-PI)
• NCCAM/NIH Dates 4/1/07-3/31/10
• Use of Energy Biofield Therapy for the Treatment
  of Cancer and GVHD.
• Not Funded
• R21 HL085768 (Courchesne Welniak, co-PI)
• NHLBI/NIH
• Antifungal Activity of Amiodarones after Bone
  Marrow Transplantation.

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Strategy for Funding Applications
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Target Funding Agencies

• NIH Funding Organizations
• National Heart, Lung and Blood Institute
• National Cancer Institute
• Private Organizations
• Leukemia and Lymphoma Society
• American Cancer Society
• Damon Runyon Cancer Research Foundation
• Peer-Reviewed Medical Research Program (DOD)

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Timeline

• Fall/Winter 2006
• Submit manuscript with key findings
• Submit application for funding from private
  agencies.
• Summer 2007
• Prepare and submit RO1 application.

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Conclusions

• Benefits from NV INBRE
• Established independent laboratory
• Provides core services that are not feasible in
  small laboratory.
• Flow Cytometry
• Histopathology
• Provides mentorship and networking with
  established investigators
• Attend weekly meetings with mentor and director
  of Flow Cytometry Core.

Networking Scientists and Resources to
Strengthen Biomedical Research in Nevada