We previously demonstrated that age-, disease- and therapy-associated effects on thymic function are fundamental causes of diminished T cell immune reconstitution following lymphocyte depletion. One could potentially overcome this limitation by enhancing thymic function or by enhancing the efficiency of thymic-independent pathways of immune reconstitution. We have focused our efforts on clinical development of recombinant human IL-7, a potent immunorestorative that works predominantly by enhancing thymic-independent immune reconstitution. We have also tested the hypothesis that immune reconstitution can be modulated in a favorable way by adoptive transfer of large numbers of resting T cells that have been manipulated to contain diminished numbers of suppressive cells. The first major accomplishment of this project during FY13 was a landmark publication documenting an important role for soluble IL7 receptor in modulating bioactivity of IL7. This project, focused on studies of the biology of IL7 receptor, was driven by the genetic observation that polymorphisms IL7R effect susceptibility to multiple sclerosis, but there was no understanding of the biological basis for these findings. Through these studies, we found that genetic variations in IL7R that predispose to autoimmunity results in a higher splicing rate of IL7R, and increased copies of mRNA lacking the transmembrane domain (e.g. exon 6). We thus predicted that polymorphisms predisposing to autoimmunity would be associated with higher levels of soluble IL7R. We tested this hypothesis using cohorts of healthy and MS-afflicted patients, and found that autoimmunity predisposing polymorphisms led to higher circulating levels of soluble IL7Ra. Furthermore, when soluble IL7Ra was added to cultures containing rhIL7, bioactivity of rhIL7 was diminished in the short-term but increased over the longer term (e.g. days), due to diminished IL7 consumption in the presence of soluble IL7Ra. We also observed qualitative differences in IL7 signaling in the presence of soluble IL7Ra, with diminished levels of SOCS and Fas induced by IL7 signaling in this context. Both of the molecules are negative regulators of T cell function, consistent with a model where higher levels of soluble IL7Ra leads to enhanced T cell activity, through both prolongation of the IL7 signal and diminished negative regulators. Together, this led to the hypothesis that polymorphisms in IL7R increase the risk of autoimmunity of enhancing the bioactivity of IL-7. Using clinical samples from healthy donors and from patient with multiple sclerosis, we confirmd that soluble IL7R levels are tightly regulated by genetic polymorphisms in IL7Ra and that in patients with multiple sclerosis, IL-7 levels themselves are also modulated by polymorphisms in IL7Ra. Finally, we observed in a mouse model of autoimmune encephalitis, that co-administration of sIL7Ra significantly increases the potency of IL-7 induced disease, which provided a clear basis for implicating soluble IL7Ra levels in susceptibility to multiple sclerosis. These studies provide novel, fundamental insights implicating an essential role for IL7R in modulating IL7 bioactivity in vivo, and elucidate a clear basis to explain how genetic polymorphisms increase susceptibility to multiple sclerosis. They are lead to the prediction that co-treatment with IL7 plus sIL7Ra will enhance the potency of IL7 as a therapeutic agent. Finally, these data reveal that under normal circumstances, humans have a profound molar excess of soluble IL7Ra to IL7 itself. This is fundamentally different that the relationship between soluble receptor found for IL2 and IL15 and likely explain the prolonged half-life of IL7 when used as a therapeutic agent. The second major accomplishment of this project in FY13 was completion of the first two cohorts of a clinical trial wherein rhIL7 was administered to a large cohort of children with cancer after primary front line therapy. This represents the only clinical experience with rhIL7 in children thus far. We demonstrated the agent to be safe and highly effective at increasing the pace and degree of immune reconstitution following intensive chemotherapy for cancer. We also conducted state-of-the-art studies using next generation sequencing to enumerate the T cell receptor repertoire of rhIL7 vs non-IL7 treated patients. These studies demonstrate diversification of the repertoire with recombinant human interleukin-7, confirming what was previously postulated using approaches of much lower resolution. Furthermore, as discussed in Project III, exciting clinical results suggest that this approach may improve survival in this high-risk population.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Investigator-Initiated Intramural Research Projects (ZIA)
Project #
1ZIABC011069-06
Application #
8763333
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
6
Fiscal Year
2013
Total Cost
$513,935
Indirect Cost
Name
National Cancer Institute Division of Basic Sciences
Department
Type
DUNS #
City
State
Country
Zip Code
Guimond, Martin; Freud, Aharon G; Mao, Hsiaoyin C et al. (2010) In vivo role of Flt3 ligand and dendritic cells in NK cell homeostasis. J Immunol 184:2769-75
Reddy, Pavan; Arora, Mukta; Guimond, Martin et al. (2009) GVHD: a continuing barrier to the safety of allogeneic transplantation. Biol Blood Marrow Transplant 15:162-8
Guimond, Martin; Veenstra, Rachelle G; Grindler, David J et al. (2009) Interleukin 7 signaling in dendritic cells regulates the homeostatic proliferation and niche size of CD4+ T cells. Nat Immunol 10:149-57
Cui, Yongzhi; Zhang, Hua; Meadors, Joanna et al. (2009) Harnessing the physiology of lymphopenia to support adoptive immunotherapy in lymphoreplete hosts. Blood 114:3831-40
Capitini, C M; Chisti, A A; Mackall, C L (2009) Modulating T-cell homeostasis with IL-7: preclinical and clinical studies. J Intern Med 266:141-53
Sportes, Claude; Hakim, Frances T; Memon, Sarfraz A et al. (2008) Administration of rhIL-7 in humans increases in vivo TCR repertoire diversity by preferential expansion of naive T cell subsets. J Exp Med 205:1701-14
Guimond, Martin; Leonard, Warren J; Spolski, Rosanne et al. (2008) Thymic stromal lymphopoietin is not necessary or sufficient to mediate the thymopoietic effects of keratinocyte growth factor. Blood 111:969-70
Dean, Robert M; Fry, Terry; Mackall, Crystal et al. (2008) Association of serum interleukin-7 levels with the development of acute graft-versus-host disease. J Clin Oncol 26:5735-41