A bioengineering method for modeling alveolar Rhabdomyosarcoma and assessing chemotherapy responses

Abstract

Rhabdomyosarcoma (RMS) is the most common pediatric soft-tissue malignant tumor. Treatment of RMS usually includes primary tumor resection along with systemic chemotherapy. Two-dimensional (2D) cell culture systems and animal models have been extensively used for investigating the potential efficacy of new RMS treatments. However, RMS cells behave differently in 2D culture than in vivo, which has recently inspired the adoption of three-dimensional (3D) culture environments. In the current paper, we will describe the detailed methodology we have developed for fabricating a 3D engineered model to study alveolar RMS (ARMS) in vitro. This model consists of a thermally cross-linked collagen disk laden with RMS cells that mimics the structural and bio-chemical aspects of the tumor extracellular matrix (ECM). This process is highly reproducible and produces a 3D engineered model that can be used to analyze the cytotoxicity and autophagy induction of drugs on ARMS cells. The most improtant bullet points are as following: We fabricated 3D model of ARMS. The current ARMS 3D model can be used for screening of chemotherapy drugs. We developed methods to detect apoptosis and autophagy in ARMS 3D model to detect the mechansims of chemotherapy agents.

Potential role of TGFΒ and autophagy in early cerebellum development

“…Autophagy, as a highly conserved lysosomal degradation and clearance pathway, is required for proper proliferation/migration, differentiation, development, remodeling, and intracellular refreshment [ [59] , [60] , [61] ]. An autophagosome marker, namely microtubule-associated protein light chain 3 beta (LC3β) contributes to the generation or degradation of autophagosomes [ [62] , [63] , [64] ]. LC3β comprised of LC3β-I and LC3β-II with a molecular weight of 16 kDa and 14 kDa, respectively, are detected in Western blot.…”

Assessment of Stiffness-Dependent Autophagosome Formation and Apoptosis in Embryonal Rhabdomyosarcoma Tumor Cells

“…Although tumor stiffness can be monitored in in vivo , it is hard to generate tumor with defined stiffness in animal models. The synthesis of hydrogels and their application in tissue engineering has made it possible to develop 3D cell cultures similar to the tumor microenvironment (20,21). The mechanical properties provided by these hydrogels regulate intracellular signaling pathways and studies similar to natural tissues can be completed with such approaches (22).…”

Regulation of Autophagy via Carbohydrate and Lipid Metabolism in Cancer

“…Autophagy is usually part of the response to chemotherapy agents in tumor cells and it can increase or decrease chemotherapy-induced apoptosis [ 28 , 29 ]. This response is dependent on the type of tumour cells and chemotherapy agents used [ 29 , 30 , 31 , 32 , 33 ]. On the other hand, autophagy-mediated cell death could also be involved in response to chemotherapy agents.…”