Tumor Immune MicroEnvironment Facility (Scientific Core)

Division Of Basic Sciences - Nci

Investigators

Linked publications & trials

Abstract

Goals and objectives: The tumor microenvironment plays a critical role in cancer development, progression, and control. The molecular and cellular nature of the tumor immune microenvironment influences disease outcome by altering the balance of suppressive versus cytotoxic responses in the vicinity of the tumor. Our focus is to study the tumor immune microenvironment (TIME) in preclinical and clinical samples from patients treated with immunotherapeutic agents in solo or in combination with other cures such as Chemotherapy or targeted therapy. In our facility, we are developing novel multiplex immunofluorescence techniques to identify immune cells, tumor cells, cytokines, etc. based on their protein expression or RNA transcription. Our studies will provide an immunological signature to provide insight into the complex biology involved immunologically mediated tumor killing or resistance to that killing. These could also be used to predict tumor progression or regression and may reveal new targets for immunotherapies for cancer patients. Project summary: We added 3 more patients to have a total of 13 patients having local advanced prostate cancer and undergoing radical prostatectomy (RP). These patients were treated with neoadjuvant PROSTVAC and Nivolumab (NCT 02933255). We investigated the immune modulation in TIME after treatment using multiplex immunofluorescence (Opal Method). FFPE sections from matched pre-treated prostate biopsies and post-treated RP samples were stained with our validated T cell panel1. Combination immunotherapy significantly increased CD4+ T cells and CD8+ T cells densities in the invasive margin, intratumoral and the benign compartments. 91% and 83% patients showed more than 2Xincrease of CD4 and CD8 T cells in the TIME, respectively, in at least one of the three compartments, showing more effect that Prostvac alone. Increased proliferative indices in CD4+ and CD8+ T cells were also seen after treatment. Tregs were present in low frequencies in TIME (average of 10/mm2) with no significant changes. Moreover, a significant drop in tumor cell Ki67 after treatment suggests that the combination may control tumor growth. The combination of Neoadjuvant Prostvac and nivolumab was associated with increased immune cell infiltration in a cohort of early prostate cancer patient. This work was submitted for a poster presentation at SITC 2023, and approval is still pending. Bone metastasis is closely related to the survival rate of cancer patients and their quality of life. In the Bone metastatic site, the tumor microenvironment plays a critical role in cancer development and progression by the interaction of immune cells and cancer cells. To characterize the landscape of immune cells infiltrating the prostate cancer Bone metastasis site, we applied 3 validated Immunofluorescence multiplex panels (lymphocyte, macrophage, and Myeloid derived suppressor cell) on 13 tumor tissue samples taken from 9 patients with castration resistant prostate cancer. These samples were provided by Thomas Jefferson University Hospital. We have finished the panels staining and analysis for all samples. The analysis is in the validation process. This work was added in preliminary studies for the PCF tactile grant that was accepted for funding in 2023. In our facility we are continuously developing new multiplex immuno-fluorescence panels to investigate more the tumor microenvironment and study more in depth its components. We have a great interest in studying the role of Natural killer cells in prostate cancer. For this purpose, we are developing a new panel that includes CD16, CD56, PDL1, CD8, Granzyme B, CK and DAPI. We optimized these antibodies in IHC and single IF and we are at the step of including them in 1 panel. This panel will also serve to look at the cytotoxic CD8 T cells and NKs in other clinical samples. To look at the activation of TGF Beta pathway in clinical tumor samples and at the impact of Bintrafusp alfa on its regulation, we are developing a multiplex panel that includes p-smad2, p-smad3, Twist, p21, E-cadherin, cytokeratin and DAPI. A great progress is made to develop this panel. In collaboration with Dr Nyall London and Riley Larkin (MSRP student), and to look at the HLA classes 1 and 2 and their interaction with CD4 and CD8 T cells in olfactory Neuroblastoma. We developed and validated a multiplex panel that includes: HLA1, HLA-DR, CD4, CD8, Ki67, Synaptophysin and DAPI. We applied this panel on an olfactory neuroblastoma Tissue micro array. Data on these cells densities and interaction was generated and presented at the AACR 2023 meeting. in collaborating with Dr Kazusa Ishii, and in the context of developing a new cell therapy TCR anti Brachyury. The expression of Brachyury in the brain tissue was to explore since in the literature there is a mixed information about this protein being expressed in the brain and in the Purkinjie cells. We used 3 antibodies anti-Brachyury from 3 different sources. For antibodies optimization we used Formalin fixed paraffin embedded cell lines: a chordoma cell line (UM-chorm1) as positive control and K562A2 as a negative cell line. The 1st and 3rd antibodies we tried showed accurate signal in the cell lines, unlike the 2nd Antibody. The Brain tissue was completely negative for Brachyury using these 2 antibodies as well. The next step will be to try these antibodies on more Normal brain samples to confirm that Brachyury is not expressed in the normal brain and therefore a TCR anti Brachyury can be developed. We are developing new IF-multiplex panels for mouse tissues to support the preclinical team at the center or immuno-oncology (CIO). The preclinical team is treating mice bearing different tumor models with different drugs in solo and in combination. To look at the effect of these drugs on TIME and correlate we are developing 6 multiplex panels using IF, each of these panels include 6 markers and DAPI. In collaboration with Dr Ling Zhang, we looked at the impact of OT1 cells and OT1 cells+IL12 in melanoma murine model using our murine lymphocyte panel that includes: CD4, CD8, Granzyme B, CD45, CD98 and DAPI. Additionally, this panel was used the look at the effect of Entinostat, PDS0101, NHS-Il12 in all combination on TC1 tumors in collaboration with Dr Caroline Jochems. Another aspect of our work is to provide material to AI teams. We are continuing our collaboration with Dr George Zeki from FNL and Dr Faisal Mahmood from Harvard Medical School and the Division of Computational Pathology at the Brigham and Women's Hospital. We started a collaboration with Dr Eytan Ruppin and his team from NCI and we provided scans of H&Es from clinical tumor samples. in this study they tested their machine learning method: ENLIGT-DeepPT on our samples to predict patients response to Bintrafusp alfa. This work was presented in ASCO 2023.