Mimicking the complex intricacies of the extra cellular matrix including 3D configurations and aligned fibrous structures were traditionally perused for producing cartilage tissue from stem cells. This study shows that human adipose derived mesenchymal stem cells (hADMSCs) establishes significant chondrogenic differentiation and may generate quality cartilage when cultured on 2D and randomly oriented fibrinogen/poly‐lactic acid nanofibers compared to 3D sandwich‐like environments. The adhering cells show well‐developed focal adhesion complexes and actin cytoskeleton arrangements confirming the proper cellular interaction with either random or aligned nanofibers. However, quantitative reverse transcription‐polymerase chain reaction analysis for Collagen 2 and Collagen 10 genes expression confirms favorable chondrogenic response of hADMSCs on random nanofibers and shows substantially higher efficacy of their differentiation in 2D configuration versus 3D constructs. These findings introduce a new direction for cartilage tissue engineering through providing a simple platform for the routine generation of transplantable stem cells derived articular cartilage replacement that might improve joint function.
BACKGROUND: Colorectal cancer (CRC) is the second most common solid organ cancer. Immunotherapy has recently emerged as a promising treatment for the MSI-H/dMMR metastatic CRC. However, MSS patients still cannot benefit from this therapy. STC therapeutic vaccine (Brenus Pharma) is composed of selected tumor cell lines, stimulated to overexpress TAA/TSA and neoantigens including resistant factors that are further haptenized to form immunogenic hapten-protein complex. This educates the immune system to recognize and target the patient’s tumor cells expressing the same resistance factors. We report results of the transcriptomic changes induced by the different treatment and culture conditions on each cell line; highlighting the potential of this three cell lines vaccine to sensitize immunotherapy-resistant tumors to anti-PD1 therapy. STUDY DESIGN and METHODS: In this study, three cell lines have been cultured: HCT116, HT29, and LoVo. Each of these cell lines has been subjected to four growth conditions: RCB) the parental cell line cultured in standard medium 10% SVF or MCB) 2% SVF; DSA) MCB condition plus low-dose irradiation and heat shock or DSB) cultured with chemotherapy exposure. STC-1010 vaccine (DP) is the mix of the haptenized DSA and DSB culture conditions of these three cell lines. Total RNA of all those samples have been extracted and sequenced on Illumina NextSeq 500. Sequences have been aligned against the GRCh38 reference genome using Kallisto v0.46.1 software. Differential abundance and enrichment analyses have been performed with DESeq2 v1.34.0 and clusterProfiler v4.2.0 R packages, respectively. RESULTS: A first overview of the whole-transcriptome shows that each cell line has its very own profile and that the DP condition reflects a balanced mixture of the three transcriptomes. HT29-DSA condition displays 1048 differentially expressed genes (DEG) compared to MCB condition and altered 59 pathways, whereas the HT29-DSB condition shows 1288 DEG and altered 46 pathways. Both conditions show an overexpression of DNA repair pathways. LoVo-DSA condition displays 1044 DEG compared to MCB condition and altered 59 pathways, especially DNA repair pathways. HT29-DSB condition shows 1791 DEG and altered 130 pathways, mostly impacting the tumor immunity. HCT116-DSA condition displays 407 DEG compared to MCB condition and altered 72 pathways, especially DNA repair pathways. HCT116-DSB condition shows 1824 DEG and altered 51 pathways. Both conditions show an overexpression of pathways involved in tumor immunity. CONCLUSION: Brenus STC vaccine based on physical and chemical stimulations and haptenization reveals a promising potential to sensitize MSS CRC immunotherapy resistant tumors to immune checkpoint inhibition therapy. Further studies are ongoing to profile and analyze the proteosurfaceome of each cell line. Citation Format: Corentin Richard, Sandy Chevrier, Lionel Chalus, Jeremy Forget, Benoît Pinteur, Paul Bravetti, Romain Boidot. Sensitizing potential of a novel stimulated and haptenized vaccine in immunotherapy resistant metastatic colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3566.
BACKGROUND: Metastatic colorectal cancer (mCRC) is one of the major causes of death worldwide. The relative therapeutic resistance of these populations to I/O drives the need for new treatment. STC-1010 (Brenus Pharma) therapeutic vaccine is developed by tumor cells stimulation to induce overexpression of TAA and neoantigens to mimic mCRC resistant cancer cells allowing education of the immune system to target the patient’s tumor cells harboring the same resistance factors. We report results of a study evaluating efficacy and safety of a 3 cell lines-based product namely “3CL-SH” (made of CT26, CMT93 and LTPA) physical stimulated (S=irradiation and heat shock) and haptenized (H) with immunostimulant in an anti-PD1 resistant MC38 colorectal tumor model grafted with its TME. METHODS: Female C57BL/6 mice bearing MC38 tumor model grown in vivo under anti-PD1 selection pressure (anti-PD1; 12.5 mg/kg; qw) to obtain MC38 resistant to anti-PD1 treatment (MC38 antiPD1R). Two sampled tumors from these mice were cut into fragment of 2 mm2 and reimplanted in the right flank of naïve syngenic mice. 5 groups (15 mice per group) were allocated to: G1) Control group: Isotype/placebo/vehicle G2) Anti-PD1/placebo/vehicule G3) Anti PD1/placebo-GM-CSF/endoxan G4) Isotype/3CL-SH-GM-CSF/endoxan G5) Anti-PD1/3CL-SH-GM-CSF/endoxan. Endoxan, vehicule and antibodies were administered by IP, vaccine or placebo subcutaneously. Overall survival (OS) and tumour growth (TG) were recorded until 1600 mm3 or safety endpoint. 5 mice per group were euthanized and sampled for immunophenotyping. In parallel, we conducted the identification and relative quantification of proteins expressed in 3CL-SH by LC-MS/MS and flow cytometry. RESULTS: Flow cytometry analysis shows that HSP-70 expression was significantly upregulated in each of the three cell lines of the 3CL-SH compared to untreated cells. LC-MS/MS analysis demonstrated that cell lines of 3CL-SH display overexpression of membrane proteins of interest such as Lamp 1, MDR1A/B and Fas Receptor. The group treated with 3CL-SH significantly improves the survival of mice bearing MC38PD1R tumor model vs the control group (Log-rank Mantel-Cox test, pvalue=0.0368) and we observed a non-benefit of the combination of 3CL-SH and anti-PD1 drug in concomitant injections. The immunophenotyping data show statistical increase in the infiltration of anti-tumor populations (CD8+ T cells, M1) in the tumors treated with 3CL-SH but also an increase in M2. No side effect or inflammatory reaction towards the 3CL-SH is evidenced. CONCLUSION: The 3CL-SH treatment presents a promising antitumor efficacy on the aggressive MC38 PD1R model, resulting in a significative gain of survival and anti-tumoral changes in the immune infiltrate of treated animals. More studies are needed to evaluate the benefit of a combination with ICI. Citation Format: Céline Gongora, Elsa Kress, Claudine Vermot-Desroches, Lionel Chalus, Jeremy Forget, Benoît Pinteur, Paul Bravetti, Charles Dumontet, Antoîne Italiano. Antitumor activity of a novel allogeneic colorectal cancer vaccine in C57BL/6 mice bearing MC38 anti-PD1 resistant colon carcinoma syngeneic model with TME [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2051.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.