Thesis
Netrin-G1 enhances in desmoplastic fibroblasts enhances interactions with pancreatic cancer cells
Master of Science (M.S.), Drexel University
Jul 2016
DOI:
https://doi.org/10.17918/etd-7164
Abstract
Pancreatic Ductal Adenocarcinoma (PDAC) is amongst the most lethal forms of cancer encountered in patients. One of the defining characteristics of this neoplasia is a large desmoplastic stromal component that includes a highly organized fibrous extracellular matrix (ECM). Previously our lab compared ECM producing normal fibroblasts to tumor associated fibroblasts (TAFs) via mRNA microarray and noticed an interesting increase in the expression of a splice variant isoform of the synapse stabilizer Netrin G1 (NTNG1). Utilizing an in vitro three dimensional culturing system, produced by patient derived TAFs, we first validated isoform expression in TAFs by western blot, immunofluorescence (in vitro and in vivo) as well as via Reverse Transcriptase Polymerase Chain Reaction. We next examined the isoform expression of NTNG1 in the presence of either fibroblastic activator Transforming Growth Factor Beta (TGF-[beta]) or its inhibitor. Our results determined that NTNG1 overexpression (compared to normal) is independent of TGF-[beta] functions such as ECM parallel fiber orientation. The corresponding tumoral receptor Netrin G1 Ligand 1 (NGL1) was also found to be highly expressed in transformed PDAC cells. CRISPR/CAS9 knockouts of NTNG1 in TAFs and of NGL1 in PDACs were used for functional analyses. PDAC spheroid spread on three dimensional TAF cultures and live imaging of co-cultures of WT and/or knockout PDACs and TAFs demonstrated that ligand-receptor expressions are necessary for stable tumoral-stromal interactions. We speculate this function to be important for stroma supported pancreatic tumor growth, whereby stroma provides nutritional tumoral support (i.e., in the absence of angiogenesis). In conclusion, we believe disruption of this interaction has further implications in pro-tumorigenic stromal regulation.
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Details
- Title
- Netrin-G1 enhances in desmoplastic fibroblasts enhances interactions with pancreatic cancer cells
- Creators
- Dustin Rollins - DU
- Contributors
- Edna Cukierman (Advisor) - Drexel University (1970-)
- Awarding Institution
- Drexel University
- Degree Awarded
- Master of Science (M.S.)
- Publisher
- Drexel University; Philadelphia, Pennsylvania
- Resource Type
- Thesis
- Language
- English
- Academic Unit
- Medicine (Graduate); College of Medicine; Hematology and Oncology; Drexel University
- Other Identifier
- 7164; 991014632932504721