Transcriptome analysis of GVHD reveals aurora kinase A as a targetable pathway for disease prevention

Abstract

Graft versus host disease (GVHD) is the most common complication of hematopoietic stem cell transplant (HCT). However, our understanding of the molecular pathways that cause this disease remains incomplete, leading to inadequate treatment strategies. To address this, we measured the gene expression profile of non-human primate (NHP) T cells during acute GVHD. Utilizing microarray technology, we measured the expression profiles of CD3+ T cells from five cohorts: allogeneic transplant recipients receiving 1) no immunoprophylaxis (No Rx); 2) sirolimus monotherapy (Siro); 3) tacrolimus-methotrexate (Tac-Mtx); as well as 4) autologous transplant recipients (Auto) and 5) healthy controls (HC). This comparison allowed us to identify transcriptomic signatures specific for allo-reactive T cells and determine the impact of both mTOR and calcineurin inhibition on GVHD-mediated pathway dysregulation. We found that the transcriptional profile of unprophylaxed GVHD was characterized by significant perturbation of pathways regulating T cell proliferation, effector function and cytokine synthesis. Within these pathways we discovered potentially druggable targets not previously implicated in GVHD, prominently including aurora kinase A (AURKA). Utilizing a murine GVHD model, we demonstrated that pharmacologic inhibition of AURKA could improve survival. Moreover, we found enrichment of AURKA transcripts both in allo-proliferating T cells and in sorted T cells from patients with clinical GVHD. These data provide a comprehensive elucidation of the T cell transcriptome in primate acute GVHD. This transcriptome enables a systems-based approach to the identification of targets for disease control, many of which are immediately amenable for clinical evaluation. The first such target identified, AURKA should now be considered a lead target for prevention of GVHD, which, given the many available AURKA inhibitors in clinical development, could be quickly deployed for the prevention of GVHD.

Systems analysis uncovers inflammatory Th/Tc17-driven modules during acute GVHD in monkey and human T cells

“…In accordance with our previously published results, 6 visualization of the top 25 gene sets in the hyperacute cohort using constellation mapping 10,11 revealed that proliferative signals were a dominant theme in the T-cell transcriptional profile of hyperacute GVHD ( Figure 3C), with 21 of 25 top correlating gene sets describing proliferative programing, and 15 of these demonstrating significant sharing of enriched transcripts (depicted as red circles in Figure 3C). Similar to our previous work, GSEA revealed that Th/Tc1 transcripts were overrepresented in animals with hyperacute GVHD compared with T cells purified from both healthy controls, and from recipients with breakthrough acute GVHD 12 ( Figure 3D).…”

“…We have previously reported the flow cytometric 4 and transcriptional attributes 6 of T cells causing GVHD in the absence of immunosuppression ("No Rx"). These attributes include a highly proliferative T-cell program, characterized by multiple markers of Th/Tc1 skewing and expression of granzymes and interferon g (IFNg).…”

“…[4][5][6] To discover the transcriptional networks that drive GVHD during clinically relevant immunoprophylaxis, we have now mapped the T-cell dysregulation that occurs in the setting of a variety of immunoprophylactic settings. We find that 2 signatures predominate: (1) a highly proliferative, cytotoxic signature that occurs during hyperacute GVHD and (2) the much more complex immune signature of breakthrough acute GVHD, which retains some T helper (Th)/T cytotoxic (Tc)1 elements, but which is predominated by an inflammatory Th/Tc17-skewed and apoptosis-resistant T-cell profile.…”

Defining success with cellular therapeutics: the current landscape for clinical end point and toxicity analysis

“…This work has been driven by detailed flow cytometric evaluation of the T-and B-cell subpopulations driving both acute and chronic disease, [86][87][88][89][90][91][92][93][94][95][96][97][98][99][100][101][102] as well as increasing use of systems-based transcriptomic approaches to identifying the pathways associated with GVHD, both in preclinical models and clinical samples. 50, [103][104][105][106][107] This work is identifying a new cohort of targetable pathways for GVHD control, many of which are amenable for clinical translation.…”

Therapeutic targets and emerging treatment options in gastrointestinal acute graft-versus-host disease

“…In experimental murine GVHD inhibition of the aurora A kinase pathway with a commercially available selective, reversible small-molecule inhibitor (MLN8237, alisertib) mitigated clinical severity and improved survival. [143] Human clinical trials have not yet been conducted.…”