10 0ctober 2018. Room 504, main building campus A of chongqing university.

Prof. Sean R.R. Hall

University of Bern, Switzerland

Interrogating The Tumor Microenvironment In Resectable Non-Small Cell Lung Cancer

There are three immune exam pathways and cancer immunotherapy in solid
epithelial tumors. First, antibodies that focus on CTLA4, PD-1 and PD-1 / L1 are
clinically effective in several types of tumors including NSCLC. Second, tumor burden,
tumor immunogenicity (mutation load/neoantigen load/presentation machine) and degree
of immune suppression in the tumor microenvironment. The third is Refractoriness and
acquiring tumor resistance.

When tumors grow, they obtain mutations, some of which create neoantigen that
affects the patient's response to immune checkpoint inhibitors. The impact of neoantigen
intratumor heterogeneity (ITH) on antitumor immunity can be explored. Through an
integrated analysis of ITH and neoantigen load, the relationship between clonal
neoantigen load and overall survival in primary pulmonary adenocarcinoma.
Lymphocytes that infiltrate neoantigen clone CD8 + tumors were identified in early-stage
non-small cell lung cancer and showed high PD-1 levels. Sensitivity to PD-1 and CTLA-
4 blockade in patients with advanced NSCLC and melanoma is enhanced in enriched
tumors for neoantigen clones. T cells that recognize clone neoantigen are detected in
patients with long-lasting clinical benefits. Subclonal neoantigens induced cytotoxic
chemotherapy, contributing to increased mutation load, were enriched in poor
respondents. These data suggest that neoantigens heterogeneity can affect immune
surveillance and support the development of therapies targeting neoantigen clones.

The immune landscape of cancer includes the composition and function of immune
cells across different tumor types, immune types of epithelial tumors, tumor
microenvironment or TME. 4 ways to interrogate tumor microenvironment or multiplex
analysis are 1. Cellular composition, 2. expression of regulating proteins (PD-1, PD-L1,
CD47), 3. intrinsic properties of tumor cells (clone architecture), 4. spatial planning
(TIL , stromal tumors and tumors). Tumors are spatially organized ecosystems (tumor-
immune microenvironment). Extensive and activated tumor stroma in NSCLC,
Identifying cells such as pericyte associated with tumors, CD90 + / CD73 + cells
occupying perivascular/peritumoral sites in NSCLC.

Pericyte tumor function can be measured using a physiologically relevant in vitro

system with a microfluidic chip, pericyte function tumor using perfusable microvessels,
microvessels with NSCL-derived pericytes. Cell immunomodulatory function derived
from pericyte tumors. Cells derived from pericyte tumors secrete factors that promote
tumors. Pericytes tumors suppress T cell activation by contact-cell and dissolve. Reversal
of pericyte tumor immunosuppression.

Therapy antibodies that block the 1-death program (PD-1) - programmed death-
ligand 1 (PD-L1) pathways can induce strong and long-lasting responses in patients with
various cancers, including metastatic urothelial cancer. However, this response only
occurs in a subset of patients. Explaining the determinants of response and resistance is
key to improving outcomes and developing new treatment strategies. Tumors from a
large group of patients with metastatic urothelial cancer treated with anti-PD-L1 agents
(atezolizumab) and identified as the main determinant of clinical outcome. The response
to treatment is associated with the CD8 + T-effector cell phenotype and, even worse, high
neoantigen or tumor burden. Lack of response is associated with the significant
transformation of β (TGFβ) growth factor signaling in fibroblasts. This occurs especially
in patients with tumors, which shows the exclusion of CD8 + T cells from tumor
parenchyma found in fibroblast-rich and collagen peritumoral stroma; usual phenotype
among patients with urothelial metastatic cancer. Using this mouse model that
recapitulates the excluded phenotype, co-administration therapy of TGFβ-blocking and
anti-PD-L1 antibodies reduces TGFβ signaling in the cell stroma, facilitates T-cell
penetration into the tumor center, and provokes strong anti-tumor immunity and tumor
regression. The integration of these three independent biological features provides the
best basis for understanding patient outcomes in these settings and shows that TGFβ
forms a tumor microenvironment to resist anti-tumor immunity by limiting T-cell
infiltration.

Summary/TME as a target of cancer therapy is a pericyte/stroma signature

predicting tumor recurrence and mortality, to understand the dynamics and heterogeneity
in the pericyte response during tumor development stage (KP: Kras LSLG12D / WT; P53
FRT/FRT, Can we reduce pericytes immunomodulatory function tumor, Therapeutic
goals in other solid tumors such as PDAC, ovary, colorectal, and breast carcinoma.

Comparing phenotypes, transcriptome, expansion and functional signatures of

intratumoral TIL using PD-1 expression. Is there a relationship between the phenotypic
features of T cells and patient outcomes: tumor recurrence and survival, Can we use PD-1
as a marker to assess the state of functional T cells and identify new drug targets, Can we
develop models to test anti-tumor T cell immunity.

Phenotype molding of stromal cells in the lung tumor microenvironment. Cancer

cells are embedded in the tumor microenvironment (TME), a complex ecosystem of
stromal cells. By comparing with matching non-malignant lung samples, we revealed
very complex TME that deeply formed stromal cells. 52 stromal cell subtypes were
identified, including new subpopulations in cell types that were previously considered
homogeneous, and transcription factors underlying their heterogeneity. For example,
fibroblasts that express different collagen sets, endothelial cells regulate homing of
immune cells and regulated genes with immune checkpoint transcripts that already exist
and correlate with T-cell activity. By assessing marker genes for this cell subtype in bulk
RNA sequencing data from 1,572 patients, immunohistochemistry for selected markers
validated them as separate cellular entities in a series of independent lung tumors.