scholarly journals Strategies to package recombinant Adeno-Associated Virus expressing the N-terminal gasdermin domain for tumor treatment

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Yuan Lu ◽  
Wenbo He ◽  
Xin Huang ◽  
Yu He ◽  
Xiaojuan Gou ◽  
...  
Keyword(s):  
Tumor Cells ◽  
Tumor Therapy ◽  
Tumor Infiltrating Lymphocytes ◽  
Oncolytic Viruses ◽  
Tumor Treatment ◽  
Adeno Associated Virus ◽  
Robust Immune Response ◽  
Cancer Models ◽  
Infiltrating Lymphocytes ◽  
Oncolytic Effect

AbstractPyroptosis induced by the N-terminal gasdermin domain (GSDMNT) holds great potential for anti-tumor therapy. However, due to the extreme cytoxicity of GSDMNT, it is challenging to efficiently produce and deliver GSDMNT into tumor cells. Here, we report the development of two strategies to package recombinant adeno-associated virus (rAAV) expressing GSDMNT: 1) drive the expression of GSDMNT by a mammal specific promoter and package the virus in Sf9 insect cells to avoid its expression; 2) co-infect rAAV-Cre to revert and express the double-floxed inverted GSDMNT. We demonstrate that these rAAVs can induce pyroptosis and prolong survival in preclinical cancer models. The oncolytic-viruses induce pyroptosis and evoke a robust immune-response. In a glioblastoma model, rAAVs temporarily open the blood-brain barrier and recruit tumor infiltrating lymphocytes into the brain. The oncolytic effect is further improved in combination with anti-PD-L1. Together, our strategies efficiently produce and deliver GSDMNT into tumor cells and successfully induce pyroptosis, which can be exploited for anti-tumor therapy.

scholarly journals Comparing syngeneic and autochthonous models of breast cancer to identify tumor immune components that correlate with response to immunotherapy in breast cancer

2021 ◽  
Author(s):  
Jessica Castrillon Lal ◽  
Anita K. Mehta ◽  
Madeline G. Townsend ◽  
Madisson Oliwa ◽  
Eric Miller ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Tumor Cells ◽  
Myeloid Cells ◽  
Tumor Infiltrating Lymphocytes ◽  
Immune Checkpoint Blockade ◽  
Laboratory Findings ◽  
Cancer Models ◽  
Mammary Fat Pad ◽  
Infiltrating Lymphocytes

Abstract Background: The heterogeneity of the breast tumor microenvironment (TME) may contribute to the lack of durable responses to immune checkpoint blockade (ICB), however, mouse models to test this are currently lacking. Proper selection and use of preclinical models are necessary for rigorous, preclinical studies to rapidly move laboratory findings into the clinic to treat patients.Methods: We compared 3 versions of a common syngeneic and autochthonous mouse model to elucidate how tumor latency and TME heterogeneity contributes to ICB resistance. We performed comprehensive characterization of the TME using quantitative flow-cytometry and RNA expression analysis (NanoString) utilizing three distinct syngeneic breast cancer models, all derived from the MMTV-PyMT autochthonous model. A commonly used protocol was used to obtain tumor cells from MMTV-PyMT mice and 1E6, 1E5 or 1E4 cells were immediately injected into the mammary fat pad of FVB/NJ wild type mice. We then performed deep immunophenotyping and tested ICB efficacy in the 3 syngeneic models compared to the autochthonous model. Results: The 4 models had vastly different TMEs that correlated to ICB responses. We found that the number of cells used to generate syngeneic tumors significantly influences tumor latency, infiltrating leukocyte population and response to ICB. Compared to the autochthonous model, all 3 syngeneic models had significantly more tumor infiltrating lymphocytes (TILs; CD3+, CD4+, and CD8+) and higher proportions of PD-L1 positive myeloid cells, whereas the MMTV-PyMT model had the highest frequency of myeloid cells out of total leukocytes. Increased TILs correlated with response to anti-PD-L1 and anti-CTLA-4 therapy; but PD-L1expression on tumor cells or PD-1 expression of T-cells did not.Conclusions: These studies reveal that the commonly used syngeneic models have low concordance with the autochthonous model. We have identified ICB-sensitive and resistant syngeneic breast cancer models, generated from the same tumor cell inoculum, and find that only the 1E4 syngeneic model is representative of the slow growing, autochthonous model. Given the lack of benefit from ICB in breast cancer, the identification of robust murine models presented here provides the opportunity to further interrogate the TME for breast cancer treatment and provide novel insights into therapeutic combinations to overcome ICB resistance.

Tumor Infiltrating Lymphocytes in Uveal Melanoma: A Link with Clinical Behavior?

2006 ◽  
Vol 19 (1) ◽  
pp. 205873920601900 ◽  
Author(s):  
S. Staibano ◽  
M. Mascolo ◽  
F. Tranfa ◽  
G. Salvatore ◽  
C. Mignogna ◽  
...  
Keyword(s):  
T Lymphocytes ◽  
Tumor Cells ◽  
Tumor Infiltrating Lymphocytes ◽  
Primary Role ◽  
Clinical Behavior ◽  
Variable Expression ◽  
Immunological Mechanisms ◽  
Variable Extent ◽  
Formalin Fixed Paraffin Embedded ◽  
Infiltrating Lymphocytes

Experimental and clinical evidence indicate that immunological mechanisms might be important in the clinical course of uveal malignant melanoma (UMM). We analyzed the amount and phenotype of tumor infiltrating lymphocytes (TIL) and the expression of the apoptosis-inducing molecule Fas and its ligand, FasL, on tumor cells and TIL in a selected series of UMM with the aim to establish if a correlation between their expression and the clinical behavior of UMM exists. TIL phenotype and Fas/FasL expression were evaluated by immunohistochemistry in 61 cases of formalin-fixed, paraffin-embedded UMM. Results were compared with the follow-up data of patients. Most of the UMM showed a prevalence of CD8+ CD3+ T lymphocytes, or CD4+ and CD8+ cells in equal amounts. UMM showed a variable expression of FasL, ranging from 0 to > 40% of neoplastic cells. Fas was always expressed in TIL, although with a variable extent. A subgroup of UMM showed in TIL a strongly reduced or even absent expression of TCR ζ-chain, involved in activation of T-lymphocytes. This subgroup was characterized by a worse outcome. We hypothesized that an impaired cytotoxic immune response due to the loss of the ζ-chain expression plays a primary role in the biological course of UMM. Our results indicate that the overcoming of the impairment of TCR function may represent a prerequisite for the development of new therapeutic strategies for managing UMM, suggesting that elimination of tumor cells may be possible by activation of cytotoxic cells present within ocular melanomas.

Intraepithelial CD94+ tumor-infiltrating lymphocytes in a context of HLA-E overexpression as a new immune checkpoint to predict prognosis in colorectal carcinomas.

2017 ◽  
Vol 35 (15_suppl) ◽  
pp. e14592-e14592
Author(s):  
Celine Bossard ◽  
Juliette Eugene ◽  
Nicolas Jouand ◽  
Delphine Dansette ◽  
Edouard Leveque ◽  
...  
Keyword(s):  
Tumor Cells ◽  
Immune Checkpoint ◽  
Immune Escape ◽  
Tumor Infiltrating Lymphocytes ◽  
High Density ◽  
Point Of View ◽  
Mhc Class Ib ◽  
Immune Resistance ◽  
Infiltrating Lymphocytes ◽  
Inhibitory Immune Checkpoint

e14592 Background: A better understanding of the immune-modulating interactions between tumor cells and immune cells underlying the balance between immune control and immune resistance in colorectal cancer (CRC) is crucial for the design of immunotherapies. We have previously demonstrated that overexpression of the human MHC class Ib molecule - HLA-E/β2 microglobulin - by tumor cells in CRC was associated with an unfavorable prognosis, suggesting its involvement in immune escape. However, the specific receptor of HLA-E/β2m - CD94/NKG2A, inhibitory or CD94/NKG2C, activating - expressed by tumor-infiltrating lymphocytes (TIL), as well as the influence of the microsatellite status in HLA-E/β2m overexpression, remain unknown. Methods: We investigated in the primary tumor of 245 CRC patients 1) the association of HLA-E/β2m overexpression and the density of CD94+ intraepithelial TIL (IEL-TIL) with the microsatellite status, 2) the nature of CD94+ TIL and the receptor expressed - CD94/NKG2A or CD94/NKG2C - and 3) the prognostic influence of CD94+ IEL-TIL. Results: HLA-E/β2m was preferentially overexpressed in MSI compared with MSS CRC (44,6 % vs 18,4 % respectively, p = 0.0001), and significantly associated with a high density of CD94+ IEL-TIL in MSI (0,9% in HLA-E/β2m+ vs 0,2% in HLA-E/β2m– CRC, p = 0,001), and in MSS CRC (0,38% vs 0,15%, p < 0,0001). These CD94+ TIL mostly corresponded to CD8+ αβ T cells preferentially expressing the inhibitory NKG2A chain. Finally, a high density of CD94+ IEL-TIL was independently associated with a worse OS (p = 0.03). Conclusions: These results strongly suggest that HLA-E/β2m - CD94/NKG2A interactions, preferentially up-regulated in MSI CRC, represent a promising inhibitory immune checkpoint. From a clinical point of view, this inhibitory immune checkpoint could be blocked by the new anti-NKG2A monoclonal antibody.

Improved in vivo efficacy of tumor-infiltrating lymphocytes after restimulation with irradiated tumor cells in vitro

1996 ◽  
Vol 3 (6) ◽  
pp. 580-587 ◽  
Author(s):  
Ulrike L. Burger ◽  
Maximilian P. Chang ◽  
Makoto Nagoshi ◽  
Peter S. Goedegebuure ◽  
Timothy J. Eberlein
Keyword(s):  
Tumor Cells ◽  
Tumor Infiltrating Lymphocytes ◽  
In Vivo Efficacy ◽  
Infiltrating Lymphocytes

scholarly journals Spatial heterogeneity and organization of tumor mutation burden and immune infiltrates within tumors based on whole slide images correlated with patient survival in bladder cancer

2019 ◽  
Author(s):  
Hongming Xu ◽  
Sunho Park ◽  
Jean René Clemenceau ◽  
Jinhwan Choi ◽  
Nathan Radakovich ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Spatial Heterogeneity ◽  
Tumor Cells ◽  
Tumor Infiltrating Lymphocytes ◽  
The Cancer Genome Atlas ◽  
Mutation Burden ◽  
Prognostic Utility ◽  
Cancer Genome Atlas ◽  
Infiltrating Lymphocytes ◽  
Whole Slide Images

AbstractHigh-TMB (TMB-H) could result in an increased number of neoepitopes from somatic mutations expressed by a patient’s own tumor cell which can be recognized and targeted by neighboring tumor-infiltrating lymphocytes (TILs). Deeper understanding of spatial heterogeneity and organization of tumor cells and their neighboring immune infiltrates within tumors could provide new insights into tumor progression and treatment response. Here we developed and applied computational approaches using digital whole slide images (WSIs) to investigate spatial heterogeneity and organization of regions harboring TMB-H tumor cells and TILs within tumors, and its prognostic utility. In experiments using WSIs from The Cancer Genome Atlas bladder cancer (BLCA), our findings show that WSI-based approaches can reliably predict patient-level TMB status and delineate spatial TMB heterogeneity and co-organization with TILs. TMB-H patients with low spatial heterogeneity enriched with high TILs show improved overall survival indicating a prognostic role of spatial TMB and TILs information in BLCA.

Sign in / Sign up

Export Citation Format

Share Document