Increased immune infiltration and chemokine receptor expression in head and neck epithelial tumors after neoadjuvant immunotherapy with the IRX-2 regimen

Abstract

IRX-2 is an injectable cancer immunotherapy composed of cytokines purified from stimulated normal-donor peripheral blood mononuclear cells. In a phase 2a trial (n = 27), neoadjuvant IRX-2 significantly increased lymphocyte infiltration (LI) into resected head and neck tumors and was associated with changes in fibrosis and necrosis. Event-free survival was 65% at 2 years, and overall survival 65% at 5 years. Overall survival was longer for patients with LI greater versus lower than the median. This substudy of the mechanisms responsible for the increase in LI with neoadjuvant IRX-2 employed multiplex immunohistochemistry (IHC) and transcriptome analysis to interrogate matched pre- and post-treatment tumor specimens from 7 available phase 2a trial patients. Multiplex IHC showed substantial increases in CD68-expressing cells (5 patients), T-cell density (4 patients), and PDL1 mean fluorescent intensity (4 patients). Consistent with IRX-2 activation of multiple immune cells, transcriptome analysis showed mean increases in expression of genes associated with NK cells, B cells, CD4+ T cells, CD8+ T cells, and dendritic cells, but not of genes associated with neutrophils. There were increases in mean expression of genes for most immune subsets, most markedly (2- to 3-fold) for B cells and dendritic cells. Mean increases in gene expression for chemokines suggest that tumor LI may be driven in part by IRX-2-induced production of chemo-attractants. Upregulation of checkpoint genes including PDL1 and CTLA4 along with increased T-cell infiltration suggests a functional antitumor immune response such that the efficacy of IRX-2 may be enhanced by combination with immune checkpoint inhibitors.

Secreted Frizzled-Related Protein 4 (SFRP4) Is an Independent Prognostic Marker in Prostate Cancers Lacking TMPRSS2: ERG Fusions

“…While SFRP4 expression measurement alone might not be sufficiently prognostic to support clinical decision making, it is of note that studies by us and others have recently identified a good number of prognostic protein markers, such as for example AZGP1 [64], EZH2 [65], p62 [66], GSK3ß [67], PSCA [68], and PTEN [36]. Especially in the light of recent technical developments enabling a simultaneous immunohistochemical analysis of up to 6 antibodies [69], we expect, that immunohistochemical expression panels may be developed for prostate cancer prognosis assessment. SFRP4 may well become an element of such a test for ERG negative prostate cancer.…”

IRX-2 natural cytokine biologic for immunotherapy in patients with head and neck cancers

“…Successful reestablishment of antitumor functions by IRX-2 stimulates an adaptive immune response characterized by upregulation of checkpoint molecules, such as CTLA4 and PDL1. 79 …”

“…In keeping with the multiplex IHC finding of an increase in PDL1 protein expression, transcriptome analysis revealed upregulation of PDL1 gene expression after treatment with IRX-2. 79 Upregulation of CTLA4 was also noted. Increased expression of these checkpoint inhibitors is indicative of an adaptive immunoresponse triggered by successful IRX-2 activation of antitumor immunofunctions.…”

Window of opportunity trials in head and neck cancer

“…[28] Motolimod + cetuximab Berinstein et al . [23] , Wolf et al . [22] IRX In a later study, Wolf et al [22] utilized subcutaneous injections of IRX-2, a biologic composed of a mixture of purified cytokines, along with cyclophosphamide, indomethacin, and zinc in a cohort of 27 patients with HNSCC.…”

“…Of 23 evaluable subjects, 4 had an objective decrease in tumor size, although this did not constitute a true partial response by RECIST criteria. Increased lymphocytic infiltration into tumors was associated with increased response and overall survival [22,23] .…”