Different pancreatic cancer microenvironments convert iPSCs into cancer stem cells exhibiting distinct plasticity with altered gene expression of metabolic pathways

Abstract

Background Cancer stem cells (CSCs) are generated under irregular microenvironment in vivo, of which mimic is quite difficult due to the lack of enough information of the factors responsible for cancer initiation. Here, we demonstrated that mouse induced pluripotent cells (miPSCs) reprogrammed from normal embryonic fibroblasts were susceptible to the microenvironment affected by cancer cells to convert into CSCs in vivo. Methods Three different pancreatic cancer line cells, BxPC3, PANC1, and PK8 cells were mixed with miPSCs and subcutaneously injected into immunodeficient mice. Tumors were evaluated by histological analysis and cells derived from iPSCs were isolated and selected from tumors. The isolated cells were characterized for cancer stem cell characters in vitro and in vivo as well as their responses to anticancer drugs. The impact of co-injection of iPSCs with cancer cells on transcriptome and signaling pathways of iPSCs was investigated. Results The injection of miPSCs mixed with human pancreatic cancer cells into immunodeficient mice maintained the stemness of miPSCs and changed their phenotype. The miPSCs acquired CSC characteristics of tumorigenicity and self-renewal. The drug responses and the metastatic ability of CSCs converted from miPSCs varied depending on the microenvironment of cancer cells. Interestingly, transcriptome profiles of these cells indicated that the pathways related with aggressiveness and energy production were upregulated from the levels of miPSCs. Conclusions Our result suggests that cancer-inducing microenvironment in vivo could rewire the cell signaling and metabolic pathways to convert normal stem cells into CSCs.

GSK-3α/β and MEK inhibitors assist the microenvironment of tumor initiation

“…We demonstrated that CM from different cells of human cancer cell lines was rich in growth factors and cytokines that could convert mouse iPSCs into CSCs (Afify et al 2020 ; Calle et al 2016 ; Chen et al 2012 ; Hassan et al 2022 ). Further, we showed that microenvironment of pancreatic cancer cell line BxPC3 cells converted mouse iPSCs into aggressive CSCs (Hassan et al 2022 ). In this context, microenvironments of different pancreatic cancer cell line cells could convert mouse iPSCs into different CSCs with different plasticity.…”

“…The hiPSCs in day 0 and after four weeks of conversion were cultured on gelatin coated dishes in DMEM media (Wako, Japan) with 15% FBS. The primary cultures of cells from tumors were prepared as same as described previously (Hassan et al 2022 ). After digestion, 3 ml of StemFit Basic 03 medium with 10% FBS was added to stop digestion.…”

“…Mice were sacrificed after 5 weeks of injections and the tumors were excised. Details for animal experiments were in our previous report (Hassan et al 2022 ). For serial transplantation of tumor tissues, the 4-week-old Balb/c nu/nu female mice (Charles River, Japan) were anesthetized for surgery.…”

“…Total RNA was extracted with TRIzol RNA isolation reagents (Life Technologies, CA, USA) and treated with DNase I (Promega, WI). Samples were subjected to sequencing with Novaseq6000 (Illumina, CA) and the obtained data were analyzed with Galaxy <usegalaxy.org> as descripted previously (Hassan et al 2022 ). Briefly, differential expressed genes were identified with Cuffdiff tool and used as inputs to detect enriched pathways from Kyoto Encyclopedia of Genes and Genomes (KEGG) database using DAVID Bioinformatics Resources (Huang et al 2009a , b ).…”

ALDOC- and ENO2- driven glucose metabolism sustains 3D tumor spheroids growth regardless of nutrient environmental conditions: a multi-omics analysis

“…On the other hand, both the presence and the abundance of nutrients within the local microenvironment may determine the metabolic phenotype that cancer cells adopt to accomplish each stage of the metastatic process. Indeed, ECM detachment causes defective glucose utilization, reduces pentose phosphate pathway (PPP), diminishes adenosine triphosphate (ATP) production, and increased ROS generation [13][14][15]. Despite advances in this field during the last decade, nutrient demands and related mechanisms that sustain the survival of cancer cells following ECM detachment have not been sufficiently elucidated.…”