Preventing relapse after allogeneic hematopoietic cell transplantation (alloHCT) requires novel approaches to identify and augment beneficial GVL alloimmunity. We propose that chronic lymphocytic leukemia (CLL) is the human "model disease" to achieve these goals and lessons learned focusing on CLL will improve all transplantation outcomes. Studying CLL patients enables sensitive measurement of minimal residual disease (MRD) which has emerged as the most important predictor of CLL disease free survival following alloHCT. Our innovative research proposal employs massively parallel sequencing of the B and T cell receptor genes to sensitively quantify CLL minimal residual disease MRD while identifying the unique T cell clones providing GVL benefit. We have already demonstrated the feasibility of high-throughput sequencing (HTS) the immunoglobulin heavy chain (IGH) for quantifying CLL MRD. In order to identify beneficial donor T cell clones, ex vivo CLL stimulated donor T cell clones will be isolated by flow cytometry and their unique V T cell receptors (TCR) will be determined by HTS. Following graft infusion, we will quantify these "marked" anti-CLL TCR clonotypes proliferation and persistence in relation to CLL MRD. Our second innovative application of HTS will assess the overall breadth and depth of donor T and B cell reconstitution following alloHCT. In the final aim, we will test the therapeutic benefit of human allogeneic cytokine induced killer cells (alloCIK) that were successfully translated through human phase I studies supported by our current PPG demonstrating feasibility and low GVHD risk. Overall, we hypothesize that impaired post-transplant donor immune reconstitution with limited T and B cell repertoire diversity limits beneficial GVL alloimmunity and associates with disease relapse. In project 4, we will pioneer massively parallel sequencing of B and T cell receptors to test this hypothesis in CLL patients undergoing alloHCT. We will: 1) quantify CLL MRD, 2) identify beneficial GVL T cell responses, 3) quantify overall immune reconstitution.

Public Health Relevance

Allogeneic hematopoietic cell transplantation can cure many hematologic malignancies through beneficial graft-versus-leukemia (GVL) immune responses. However, relapse remains the major cause of death. Chronic Lymphocytic leukemia (CLL) is the most common blood cancer, can be frequently cured following alloHCT via GVL benefit, and CLL MRD response predicts cure. Our novel approaches to identify and augment CLL GVL will improve all transplantation outcomes.

National Institute of Health (NIH)
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Special Emphasis Panel (ZCA1)
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Stanford University
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Sega, Emanuela I; Leveson-Gower, Dennis B; Florek, Mareike et al. (2014) Role of lymphocyte activation gene-3 (Lag-3) in conventional and regulatory T cell function in allogeneic transplantation. PLoS One 9:e86551
Hongo, D; Tang, X; Baker, J et al. (2014) Requirement for interactions of natural killer T cells and myeloid-derived suppressor cells for transplantation tolerance. Am J Transplant 14:2467-77
Florek, Mareike; Sega, Emanuela I; Leveson-Gower, Dennis B et al. (2014) Autologous apoptotic cells preceding transplantation enhance survival in lethal murine graft-versus-host models. Blood 124:1832-42
Benjamin, Jonathan; Chhabra, Saurabh; Kohrt, Holbrook E et al. (2014) Total lymphoid irradiation-antithymocyte globulin conditioning and allogeneic transplantation for patients with myelodysplastic syndromes and myeloproliferative neoplasms. Biol Blood Marrow Transplant 20:837-43
Medeiros, B C; Tian, L; Robenson, S et al. (2014) European LeukemiaNet classification intermediate risk-1 cohort is associated with poor outcomes in adults with acute myeloid leukemia undergoing allogeneic hematopoietic cell transplantation. Blood Cancer J 4:e216
Popli, Rakesh; Sahaf, Bita; Nakasone, Hideki et al. (2014) Clinical impact of H-Y alloimmunity. Immunol Res 58:249-58
Logan, Aaron C; Vashi, Nikita; Faham, Malek et al. (2014) Immunoglobulin and T cell receptor gene high-throughput sequencing quantifies minimal residual disease in acute lymphoblastic leukemia and predicts post-transplantation relapse and survival. Biol Blood Marrow Transplant 20:1307-13
Logan, A C; Zhang, B; Narasimhan, B et al. (2013) Minimal residual disease quantification using consensus primers and high-throughput IGH sequencing predicts post-transplant relapse in chronic lymphocytic leukemia. Leukemia 27:1659-65
Colonna, Lucrezia; Florek, Mareike; Leveson-Gower, Dennis B et al. (2013) IL-17 gene ablation does not impact Treg-mediated suppression of graft-versus-host disease after bone marrow transplantation. Biol Blood Marrow Transplant 19:1557-65
Shamloo, Amir; Manchandia, Milan; Ferreira, Meghaan et al. (2013) Complex chemoattractive and chemorepellent Kit signals revealed by direct imaging of murine mast cells in microfluidic gradient chambers. Integr Biol (Camb) 5:1076-85

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