Curcuphenol possesses an unusual histone deacetylase enhancing activity that counters immune escape in metastatic tumours

Curcuphenol, a common component of the culinary spices, naturally found in marine invertebrates and plants, has been identified as a novel candidate for reversing immune escape by restoring expression of the antigen presentation machinery (APM) in invasive cancers, thereby resurrecting the immune recognition of metastatic tumours. Two synthetic curcuphenol analogues, were prepared by informed design that demonstrated consistent induction of APM expression in metastatic prostate and lung carcinoma cells. Both analogues were subsequently found to possess a previously undescribed histone deacetylase (HDAC)-enhancing activity. Remarkably, the H3K27ac ChIPseq analysis of curcuphenol-treated cells reveals that the induced epigenomic marks closely resemble the changes in genome-wide pattern observed with interferon-γ, a cytokine instrumental for orchestrating innate and adaptive immunity. These observations link dietary components to modifying epigenetic programs that modulate gene expression guiding poised immunity

Studies on immune escape mechanism in cancer

Metastatic cancer is the leading cause of death in Canada. The personalized medical framework considers each patient’s genetic profile as a background to develop a specific treatment approach. Due to the success of early diagnostics, the mutational landscape of a tumour is mainly based on the sequencing data collected from early resected primary tumours, which do not necessarily reflect the mutational heterogeneity of the metastatic form of the disease and/or local recurrences underestimating tumour adaptational variations, challenging biomarker development and hindering therapeutic strategies of personalized medicine. This study has been designed to define the changes in metastatic potential of a developing tumour highlighting the immunological tumour properties, as one of emerging cancer hallmark. Microarray profiling of two separate paired cell lines of murine lung and prostate carcinomas allowed the detection of IFI44 and IL-33 as possible regulators of immunological properties within tumours. Significant down-regulation of these genes allows cancer cells with high metastatic potential to acquire capabilities to avoid destruction by the immune system via suppression of MHC-I expression. The immune-evasive phenotype allows tumour to obtain biological advantages resulting in the evasion of eradication by the immune system, which is a significant barrier for tumour growth and progression. Further study found that the overexpression of these selected genes reverses the antigen presentation deficiency in murine metastatic lung carcinoma and makes the tumour recognizable to the immune system. A parallel human study demonstrated that the expression of IL-33 is also co-regulated with HLA-I levels in human prostate cancer. Moreover, IL-33 by itself may be used as an immune prognostic biomarker for recurrence and survival in human prostate and kidney renal clear cell carcinomas. This new link in cancer biology allowed for the development a novel immunotherapeutic strategy for cancer-free survival via IL-33/ILC2 axis and the use of adoptively-transferred ILC2s. Testing this strategy on ILC2-deficient animals and animals that received ILC2s via adoptive transfer showed the importance of IL-33 and ILC2s in reducing tumour growth rate and metastatic spread to distal organs. Collectively, this thesis demonstrates a new mechanism for tumour immune escape and suggests a novel immunotherapeutic approach for anti-cancer treatment.Science, Faculty ofMicrobiology and Immunology, Department ofGraduat

A novel type-2 innate lymphoid cell-based immunotherapy for cancer

Cell-based cancer immunotherapy has achieved significant advancements, providing a source of hope for cancer patients. Notwithstanding the considerable progress in cell-based immunotherapy, the persistently low response rates and the exorbitant costs associated with their implementation still present a formidable challenge in clinical settings. In the landscape of cell-based cancer immunotherapies, an uncharted territory involves Type 2 innate lymphoid cells (ILC2s) and interleukin-33 (IL-33) which promotes ILC2 functionality, recognized for their inherent ability to enhance immune responses. Recent discoveries regarding their role in actuating cytolytic T lymphocyte responses, including curbing tumor growth rates and hindering metastasis, have added a new dimension to our understanding of the IL-33/ILC2 axis. These recent insights may hold significant promise for ILC2 cell-based immunotherapy. Nevertheless, the prospect of adoptively transferring ILC2s to confer immune protection against tumors has yet to be investigated. The present study addresses this hypothesis, revealing that ILC2s isolated from the lungs of tumor-bearing mice, and tumor infiltrating ILC2s when adoptively transferred after tumor establishment at a ratio of one ILC2 per sixty tumor cells, leads to an influx of tumor infiltrating CD4+ and CD8+ T lymphocytes as well as tumor infiltrating eosinophils resulting in a remarkable reduction in tumor growth. Moreover, we find that post-adoptive transfer of ILC2s, the number of tumor infiltrating ILC2s is inversely proportional to tumor size. Finally, we find corollaries of the IL-33/ILC2 axis enhancing the infiltration of eosinophils in human prostate carcinomas patients' expressing high levels of IL-33 versus those expressing low levels of IL-33. Our results underscore the heightened efficacy of adoptively transferred ILC2s compared to alternative approaches, revealing an approximately one hundred fifty-fold superiority on a cell-per-cell basis over CAR T-cells in the specific targeting and elimination of tumors within the same experimental model. Overall, this study demonstrates the functional significance of ILC2s in cancer immunosurveillance and provides the proof of concept of the potential utility of ILC2 cell-based cancer immunotherapies

Serum free culture for the expansion and study of type 2 innate lymphoid cells

Abstract Type 2 innate lymphoid cells (ILC2s) were discovered approximately ten years ago and their clinical relevance is gaining greater importance. However, their successful isolation from mammalian tissues and in vitro culture and expansion continues to pose challenges. This is partly due to their scarcity compared to other leukocyte populations, but also because our current knowledge of ILC2 biology is incomplete. This study is focused on ST2+ IL-25Rlo lung resident ILC2s and demonstrate for the first time a methodology allowing mouse type 2 innate lymphoid cells to be cultured, and their numbers expanded in serum-free medium supplemented with Interleukins IL-33, IL-2, IL-7 and TSLP. The procedures described methods to isolate ILC2s and support their growth for up to a week while maintaining their phenotype. During this time, they significantly expand from low to high cell concentrations. Furthermore, for the first time, sub-cultures of primary ILC2 purifications in larger 24- and 6-well plates were undertaken in order to compare their growth in other media. In culture, ILC2s had doubling times of 21 h, a growth rate of 0.032 h−1 and could be sub-cultured in early or late phases of exponential growth. These studies form the basis for expanding ILC2 populations that will facilitate the study and potential applications of these rare cells under defined, serum-free conditions

Type 2 Innate Lymphocytes Actuate Immunity Against Tumours and Limit Cancer Metastasis

Abstract Type 2 innate lymphoid cells (ILC2) potentiate immune responses, however, their role in mediating adaptive immunity in cancer has not been assessed. Here, we report that mice genetically lacking ILC2s have significantly increased tumour growth rates and conspicuously higher frequency of circulating tumour cells (CTCs) and resulting metastasis to distal organs. Our data support the model that IL-33 dependent tumour-infiltrating ILC2s are mobilized from the lungs and other tissues through chemoattraction to enter tumours, and subsequently mediate tumour immune-surveillance by cooperating with dendritic cells to promote adaptive cytolytic T cell responses. We conclude that ILC2s play a fundamental, yet hitherto undescribed role in enhancing anti-cancer immunity and controlling tumour metastasis