isolation-proximal-fluids-to-investigate-tumor-microenvironment

Isolation of Proximal Fluids to Investigate the Tumor Microenvironment of Pancreatic Adenocarcinoma

Pancreatic adenocarcinoma (PDAC) is the fourth leading cause of cancer-related death, and soon to become the second. There is an urgent need of variables ...

Humanitas Clinical and Research Center-IRCCS

University of Milan

Tumor-associated macrophages (TAMs) provide a nurturing microenvironment for metastasis and are concomitantly key determinants of the efficacy of anticancer strategies. TAM represent an extremely heterogeneous population in terms of cell morphology, functions, and tissue localization. Colorectal liver metastases (CLM) display a high heterogeneity, responsible for a wide array of clinical presentations and responsiveness to treatments. In the era of precision medicine, there is a critical need of reliable prognostic markers to improve patient stratification, and, for their predominance in metastatic tissues, TAMs are emerging as promising candidates.

Macrophages in Colorectal Cancer Liver Metastases.

Ectopic (or tertiary) lymphoid tissue develops at sites of inflammation or infection in non lymphoid organs and is associated with chronic inflammation. In colon mucosa, small lymphoid aggregates are already present in homeostatic conditions, as part of the gut-associated lymphoid tissue and play an essential role in the immune response to perturbations of the mucosal microenvironment. Despite the recognized role of inflammation in tumor progression, the presence and biological function of lymphoid tissue in cancer has been poorly investigated. We identified aggregates of lymphocytes resembling tertiary lymphoid tissue in human colorectal cancer specimens; intratumor accumulations of lymphocytes display a high degree of compartmentalization, with B and T cells, mature dendritic cells and a network of CD21+ follicular dendritic cells (FDC). We analyzed the adaptation of colon lymphoid tissue in a murine model of colitis-associated cancer (AOM/DSS). B cell follicle formation increases in the context of the chronic inflammation associated to intestinal neoplasia, in this model. A network of lymphatic and haematic vessels surrounding B cell follicles is present and includes high endothelial venules (HEV). Future task is to determine whether lymphoid tissue contributes to the persistence of the tumor-associated inflammatory reaction, rather than represent a functional immune compartment, potentially participating to the anti tumor response. 

Tertiary intratumor lymphoid tissue in colo-rectal cancer.

Macrophages are essential elements in the tumor microenvironment, where they can promote tumor growth but also influence the efficacy of anticancer strategies. In conventional therapies, chemotherapy and radiotherapy, TAMs play a dichotomous role, contributing to antitumor activity or hindering the efficacy of cytoreductive therapies. Macrophages express checkpoint ligands and are therefore targets of immunotherapy approaches based on checkpoint inhibitors. Targeted therapies with monoclonal antibodies elicit TAMs to engage in antitumor functions such as antibody-dependent phagocytosis through the activation of Fc receptors. New approaches to exploit macrophage effector functions induced by therapeutic antibodies are under investigation. Finally, strategies aimed at targeting TAM recruitment, survival and functional polarization are advancing towards the clinic. Collectively, TAM-centered strategies will hopefully complement conventional and unconventional anticancer therapies to achieve improved therapeutic benefit.

Macrophages at the crossroads of anticancer strategies.

Better understanding of pancreatic diseases, including pancreatic ductal adenocarcinoma (PDAC), is an urgent medical need, with little advances in preoperative differential diagnosis, preventing rational selection of therapeutic strategies. The clinical management of pancreatic cancer patients would benefit from the identification of variables distinctively associated with the multiplicity of pancreatic disorders. We investigated, by H nuclear magnetic resonance, the metabolomic fingerprint of pancreatic juice (the biofluid that collects pancreatic products) in 40 patients with different pancreatic diseases. Metabolic variables discriminated PDAC from other less aggressive pancreatic diseases and identified metabolic clusters of patients with distinct clinical behaviors. PDAC specimens were overtly glycolytic, with significant accumulation of lactate, which was probed as a disease-specific variable in pancreatic juice from a larger cohort of 106 patients. In human PDAC sections, high expression of the glucose transporter GLUT-1 correlated with tumor grade and a higher density of PD-1 T cells, suggesting their accumulation in glycolytic tumors. In a preclinical model, PD-1 CD8 tumor-infiltrating lymphocytes differentially infiltrated PDAC tumors obtained from cell lines with different metabolic consumption, and tumors metabolically rewired by knocking down the phosphofructokinase () gene displayed a decrease in PD-1 cell infiltration. Collectively, we introduced pancreatic juice as a valuable source of metabolic variables that could contribute to differential diagnosis. The correlation of metabolic markers with immune infiltration suggests that upfront evaluation of the metabolic profile of PDAC patients could foster the introduction of immunotherapeutic approaches for pancreatic cancer.

Metabolome of Pancreatic Juice Delineates Distinct Clinical Profiles of Pancreatic Cancer and Reveals a Link between Glucose Metabolism and PD-1 Cells.

Immunotherapy in hepatobiliary tumors: search for the missing pieces of the puzzle.

It has long been known that in vitro polarized macrophages differ in morphology. Stemming from a conventional immunohistology observation, we set out to test the hypothesis that morphology of tumor-associated macrophages (TAMs) in colorectal liver metastasis (CLM) represents a correlate of functional diversity with prognostic significance. Density and morphological metrics of TAMs were measured and correlated with clinicopathological variables. While density of TAMs did not correlate with survival of CLM patients, the cell area identified small (S-TAM) and large (L-TAM) macrophages that were associated with 5-yr disease-free survival rates of 27.8% and 0.2%, respectively (P < 0.0001). RNA sequencing of morphologically distinct macrophages identified LXR/RXR as the most enriched pathway in large macrophages, with up-regulation of genes involved in cholesterol metabolism, scavenger receptors, MERTK, and complement. In single-cell analysis of mononuclear phagocytes from CLM tissues, S-TAM and L-TAM signatures were differentially enriched in individual clusters. These results suggest that morphometric characterization can serve as a simple readout of TAM diversity with strong prognostic significance.

Macrophage morphology correlates with single-cell diversity and prognosis in colorectal liver metastasis.

The peripheral nervous system and the immune system are critically involved in the surveillance of our body, having the ability to sense the environment, recognize danger signals and orchestrate an appropriate response. Despite the fact that these two systems have been historically considered autonomous entities, a large body of evidence has shown how they interact in many homeostatic responses and how these interactions are critically involved in pathologic contexts too, including inflammation, infection and autoimmunity. In the context of cancer, where it is already known that inflammation plays a key role, the cross-regulation of immune cells and neural components is still somewhat unexplored. Detailed characterization of the mediators and pathways involved in neuro-immune interactions in cancer is expected to provide insights into the pathogenesis of disease and open new possibilities related to therapeutic strategies.

The neuro-immune axis in cancer: Relevance of the peripheral nervous system to the disease.

Duodenal adenocarcinoma (DA) is a rare yet aggressive malignancy, with increasing incidence in the last decades. Its low frequency has hampered a thorough understanding of the pathogenesis of the disease and of its biology, limiting the identification of tailored therapeutic options. A large body of evidence has clearly shown the clinical relevance of immune cells in solid tumors, correlating immune features with post-surgical prognosis. The aim of this study was to analyze the immune contexture in a cohort of duodenal adenocarcinomas surgically resected at our Institution and define its correlation with clinical variables.

Immune infiltrating cells in duodenal cancers.

Tumor tissues are populated by a multitude of macrophages, highly different in functional activity, localization and morphology. A clear contribution to disease progression has been shown in multiple cancer types, holding promise for the development of innovative macrophage-based prognostic tools. Current studies aimed at assessing the prognostic role of macrophages have documented the relevance of the macrophage population as a whole. However, dissecting the diversity of mononuclear phagocytes in tumor tissues has provided important information about the coexistence of distinct populations of macrophages with different prognostic significance. Here we summarize evidence of macrophage prognostic function in human cancer and focus on classical and modern strategies aimed at measuring macrophage features and deciphering their diversity. The wealth of new data generated will reshape our knowledge of macrophage complexity and hopefully foster the forthcoming development of these new metrics into prognostic tools as well as new therapeutic strategies.

Article	Year
Macrophages in Colorectal Cancer Liver Metastases. Cancers\| PubMed ID: 31067629	2019
Tertiary intratumor lymphoid tissue in colo-rectal cancer. Cancers\| PubMed ID: 24213222	2011
Macrophages at the crossroads of anticancer strategies. Frontiers in bioscience (Landmark edition)\| PubMed ID: 31136979	2019
Metabolome of Pancreatic Juice Delineates Distinct Clinical Profiles of Pancreatic Cancer and Reveals a Link between Glucose Metabolism and PD-1 Cells. Cancer immunology research\| PubMed ID: 32019781	2020
Immunotherapy in hepatobiliary tumors: search for the missing pieces of the puzzle. Hepatobiliary surgery and nutrition\| PubMed ID: 32142066	2020
Macrophage morphology correlates with single-cell diversity and prognosis in colorectal liver metastasis. The Journal of experimental medicine\| PubMed ID: 32785653	2020
The neuro-immune axis in cancer: Relevance of the peripheral nervous system to the disease. Immunology letters\| PubMed ID: 32827634	2020
Immune infiltrating cells in duodenal cancers. Journal of translational medicine\| PubMed ID: 32883314	2020
Prognostic significance of tumor-associated macrophages: past, present and future. Seminars in immunology\| PubMed ID: 32943279	2020

Federica Marchesi

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