Allogeneic bone marrow or hematopoietic stem cell transplantation (allo BMT) is an important therapy with curative potential for a variety of malignant and non-malignant diseases. Despite standard prophylactic regimens, graft-versus-host disease (GVHD) continues to limit the success of outcomes in allo BMT patients. Gastrointestinal (GI) GVHD is the predominant contributor to acute GVHD-related mortality. In the first funding period we hypothesized that T cell trafficking to the intestines represented a therapeutic target that could significantly reduce intestinal and systemic GVHD while sparing graft-versus-tumor and improve overall outcome. Among other results we identified the ?4?7 integrin as an excellent target for the prevention of GI GVHD and we are currently developing a clinical protocol with a neutralizing antibody. In this renewal application, we propose to continue exploring the pathophysiology of intestinal GVHD and focus on the role of the intestinal flora. Early studies in mice and in patients suggested a link between an individual's intestinal microbial flora and his/her propensity for GVHD, but the translation to the clinic (gut decontamination and/or laminar airflow protective environment) remains controversial. The availability of a novel technology, 16s ribosomal RNA (rRNA) gene sequencing, to analyze the composition of the gut flora in much greater detail has resulted in better understanding of the relationship between gut flora and its interaction with intestinal immunity and intestinal epithelil homeostasis. Our preliminary data in mouse models indicate that GVHD is associated with specific changes in the intestinal flora, including a loss of microbial diversity, dysbiosis (imbalance of intestinal flora) and the expansion of Lactobacillus or Enterobacteriaceae, which have opposite effects on GVHD. Moreover, pre-BMT antibiotic use could aggravate GVHD. Our analysis of 66 allo BMT patients shows a) a remarkable loss of microbial diversity in allo-HSCT recipients, which is associated with bacteremia and intestinal domination by Enterococci, Streptococci and Staphylococci, and b) an association between dysbiosis and GVHD. Therefore, we hypothesize that the interactions between intestinal microbiota, allogeneic donor cells, intestinal immunity and intestinal epithelial homeostasis can impact GVHD, and this can be exploited therapeutically to reduce GVHD. We propose in Aim 1 to fully characterize the intestinal microbiota during allo BMT. We will assess the effects of the conditioning and antibiotic regimens and further characterize the GVHD-associated changes (loss of microbial diversity, dysbiosis and expansion of Lactobacillus or Enterobacteriaceae). These experiments are required to define the variables that affect the intestinal flora in allo BMT recipients and provide the necessary background for the experiments in Aim 2, in which we hypothesize that manipulation of the intestinal flora and nutrition can ameliorate intestinal GVHD. We will study the effects of a) introduction of Lactobacillus on GVHD and potential mechanisms of action (suppression of Enterococcus and ??T cells), b) other potentially anti-inflammatory bacteria on GVHD (prioritizing Bifidobacterium), and c) the diet content of aryl hydrocarbon receptor ligands on the intestinal flora and GVHD. In addition, we will explore effects of murine and human intestinal flora on GVHD in germ-free and gnotobiotic mice. The investigators of this proposal are performing similar sequential analyses of the gut flora in allo BMT patients, which fosters a continuing interaction and translatability of the results. Therefore, we expect that these preclinical studies have a high likelihood to improve prophylaxis and treatment of intestinal GVHD as well as overall outcome in allo BMT patients.

Public Health Relevance

Allogeneic hematopoietic stem cell transplantation is performed to cure people of life threatening cancers (mostly of the blood) and a variety of other diseases, including auto-immune diseases. One of its major limitations is the life threatening complication of intestinal graft-versus-host disease. We will study interactions between the immune system, the intestines, and the bacteria within the intestines to develop new therapies for the prevention or treatment of intestinal graft-versus-host disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
9R01AI100288-06A1
Application #
8297808
Study Section
Cancer Immunopathology and Immunotherapy Study Section (CII)
Program Officer
Nabavi, Nasrin N
Project Start
2004-04-01
Project End
2017-06-30
Budget Start
2012-07-01
Budget End
2013-06-30
Support Year
6
Fiscal Year
2012
Total Cost
$411,525
Indirect Cost
$186,525
Name
Sloan-Kettering Institute for Cancer Research
Department
Type
DUNS #
064931884
City
New York
State
NY
Country
United States
Zip Code
10065
Tuckett, Andrea Z; Thornton, Raymond H; Shono, Yusuke et al. (2014) Image-guided intrathymic injection of multipotent stem cells supports lifelong T-cell immunity and facilitates targeted immunotherapy. Blood 123:2797-805
Geiger, Theresa L; Abt, Michael C; Gasteiger, Georg et al. (2014) Nfil3 is crucial for development of innate lymphoid cells and host protection against intestinal pathogens. J Exp Med 211:1723-31
Hubbard-Lucey, Vanessa M; Shono, Yusuke; Maurer, Katie et al. (2014) Autophagy gene Atg16L1 prevents lethal T cell alloreactivity mediated by dendritic cells. Immunity 41:579-91
Shono, Yusuke; Tuckett, Andrea Z; Ouk, Samedy et al. (2014) A small-molecule c-Rel inhibitor reduces alloactivation of T cells without compromising antitumor activity. Cancer Discov 4:578-91
Ghosh, Arnab; Holland, Amanda M; van den Brink, Marcel R M (2014) Genetically engineered donor T cells to optimize graft-versus-tumor effects across MHC barriers. Immunol Rev 257:226-36
Velardi, Enrico; Dudakov, Jarrod A; van den Brink, Marcel R M (2013) Clinical strategies to enhance thymic recovery after allogeneic hematopoietic stem cell transplantation. Immunol Lett 155:31-5
Ghosh, Arnab; Holland, Amanda M; Dogan, Yildirim et al. (2013) PLZF confers effector functions to donor T cells that preserve graft-versus-tumor effects while attenuating GVHD. Cancer Res 73:4687-96