Establishment of a tissue-engineered colon cancer model for comparative analysis of cancer cell lines.

To overcome the limitations of in vitro two-dimensional (2D) cancer models in mimicking the complexities of the native tumor milieu, three-dimensional (3D) engineered cancer models using biomimetic materials have been introduced to more closely recapitulate the key attributes of the tumor microenvironment. Specifically, for colorectal cancer (CRC), a few studies have developed 3D engineered tumor models to investigate cell-cell interactions or efficacy of anti-cancer drugs. However, recapitulation of CRC cell line phenotypic differences within a 3D engineered matrix has not been systematically investigated. Here, we developed an in vitro 3D engineered CRC (3D-eCRC) tissue model using the natural-synthetic hybrid biomaterial PEG-fibrinogen and three CRC cell lines, HCT 116, HT-29, and SW480. To better recapitulate native tumor conditions, our 3D-eCRC model supported higher cell density encapsulation (20 × 106 cells/mL) and enabled longer term maintenance (29 days) as compared to previously reported in vitro CRC models. The 3D-eCRCs formed using each cell line demonstrated line-dependent differences in cellular and tissue properties, including cellular growth and morphology, cell subpopulations, cell size, cell granularity, migration patterns, tissue growth, gene expression, and tissue stiffness. Importantly, these differences were found to be most prominent from Day 22 to Day 29, thereby indicating the importance of long-term culture of engineered CRC tissues for recapitulation and investigation of mechanistic differences and drug response. Our 3D-eCRC tissue model showed high potential for supporting future in vitro comparative studies of disease progression, metastatic mechanisms, and anti-cancer drug candidate response in a CRC cell line-dependent manner.

Consensus molecular subtype differences linking colon adenocarcinoma and obesity revealed by a cohort transcriptomic analysis.

Colorectal cancer (CRC) is the third-leading cause of cancer-related deaths in the United States and worldwide. Obesity-a worldwide public health concern-is a known risk factor for cancer including CRC. However, the mechanisms underlying the link between CRC and obesity have yet to be fully elucidated in part because of the molecular heterogeneity of CRC. We hypothesized that obesity modulates CRC in a consensus molecular subtype (CMS)-dependent manner. RNA-seq data and associated tumor and patient characteristics including body weight and height data for 232 patients were obtained from The Cancer Genomic Atlas-Colon Adenocarcinoma (TCGA-COAD) database. Tumor samples were classified into the four CMSs with the CMScaller R package; body mass index (BMI) was calculated and categorized as normal, overweight, and obese. We observed a significant difference in CMS categorization between BMI categories. Differentially expressed genes (DEGs) between obese and overweight samples and normal samples differed across the CMSs, and associated prognostic analyses indicated that the DEGs had differing associations on survival. Using Gene Set Enrichment Analysis, we found differences in Hallmark gene set enrichment between obese and overweight samples and normal samples across the CMSs. We constructed Protein-Protein Interaction networks and observed differences in obesity-regulated hub genes for each CMS. Finally, we analyzed and found differences in predicted drug sensitivity between obese and overweight samples and normal samples across the CMSs. Our findings support that obesity impacts the CRC tumor transcriptome in a CMS-specific manner. The possible associations reported here are preliminary and will require validation using in vitro and animal models to examine the CMS-dependence of the genes and pathways. Once validated the obesity-linked genes and pathways may represent new therapeutic targets to treat colon cancer in a CMS-dependent manner.

Effect of Chrysophyllum albidum fruit pulp powder on antioxidant and proinflammatory genes in non-diabetic and type 2 diabetic rats.

BACKGROUND: Diabetes mellitus (DM) is a metabolic disorder characterized by chronic hyperglycemia resulting from insulin deficiency or dysfunction. The imbalance between free radicals and antioxidants known as oxidative stress has been implicated in the pathogenesis and complications associated with DM. Chrysophyllum albidum is a seasonal fruit found to be rich in natural antioxidants. METHODS: DM was induced by high-fat diet dietary supplementation for 14 days followed by intraperitoneal injection of streptozotocin (35 mg/kg). Thirty-five experimental rats were then divided into seven groups viz.: non-diabetic control; diabetic control; metformin; diabetic and non-diabetic fed with 5 and 10% C. albidum fruit pulp powder (CAFPP). Fasting blood glucose was done with an automatic auto-analyzer and weights were monitored at three-day intervals. The expressions of Nrf2, SOD, CAT, GST, TNF-α, DPP4, and insulin were investigated using RT-PCR. Schrödinger suites was used for docking of C. albidum phytocompounds with insulin. RESULTS: Diabetic rats fed with CAFPP for thirteen days have their blood glucose lowered significantly (p < 0.05) and gained weight compared to diabetic control. CAFPP significantly (p < 0.05) up-regulated Nrf2, CAT, GST, SOD, and insulin genes expression in the diabetic group relative to diabetic control with concomitant down-regulation of TNF-α and DPP4 genes expression. Molecular docking of compounds previously characterized from C. albidum revealed that they are potent ligands of insulin receptors. CONCLUSION: The study revealed that CAFPP could be effective in the management of DM-related oxidative stress by up-regulating antioxidant and down-regulating pro-inflammatory genes expression. It also positively modulates genes associated with glucose metabolism. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s40200-021-00921-0.

African star apple fruit pulp-supplemented diet modulates fertility-related biomolecules in the testis and epididymis of high-fat diet/streptozotocin-induced diabetic rats.

This study evaluated the modulatory effect of African star apple fruit (ASAF) pulp inclusive diet on biomolecules associated with reproductive function in the testis and epididymis of high-fat diet/streptozotocin-induced diabetic male rats. The rats were divided into seven groups: control, diabetic control, diabetic rats treated with metformin, diabetic rats served with diet having 5 and 10% ASAF, respectively, and control rats served with diet containing 5%, and 10% ASAF respectively for 14 days. There were noticeable decrease in sperm parameters, reproductive hormone, glycogen, nitric oxide, total thiol, nonprotein thiol levels, and testicular 3ß- and 17ß-hydroxysteroid dehydrogenase activities and concomitant increase in cholesterol, reactive oxygen species, malondialdehyde levels, and arginase activity compared to the control. Nevertheless, ASAFs reversed all these parameters toward the control levels. Therefore, these findings suggest that ASAF pulp-supplemented diet might be an active approach in controlling male reproductive dysfunction induced by diabetes. PRACTICAL APPLICATIONS: The results suggest that ASAF pulp-supplemented diet might be an active approach in controlling male reproductive dysfunction induced by diabetes through alterations in the levels of blood glucose, glycogen, cholesterol, nitric oxide, reproductive hormones, activities of steroidogenic enzymes, arginase, and sperm characteristics as well as the antioxidant status in the testes and epididymis.