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Case Study // NOV 10 2014

Mesenchymal stem cells (MSCs) transfected with TRAIL-bearing vectors provide an antitumor therapy

Generating clinically relevant TNF-related apoptosis inducing ligand (TRAIL) engineered pancreas derived MSCs

"TRAIL-bearing expression plasmids were used for the transfection, thereby avoiding viral vectors in view of potential clinical studies. Two types of plasmids were used: non-secreting TRAIL (nsTRAIL) and secreting TRAIL (stTRAIL) as previously shown.24 The transfections were performed with TransIT®-2020 (Mirus, Madison, WI, USA) as suggested by the manufacturer."

- Moniri et al./em>

Background:

Pancreatic cancer represents 3% of all cancers, and due to the lack of curative surgeries and effective therapies, less than one in five patients survive the disease for more than one year post-diagnosis. One potential approach is to exploit the tumor-homing capacity of mesenchymal stem cells (MSCs). Specific migration of MSCs to tumors has been demonstrated in a number of clinical studies in vitro and using in vivo animal tumor models. Furthermore, engineering of MSCs to express anti-cancer genes can provide a biotherapeutic approach that can act directly and continuously at the tumor site.

In the present study, human pancreatic tumor cells were isolated from human pancreas ductal tissue, expanded ex vivo, characterized and engineered to express a known anticancer agent, tumor necrosis factor (TNF)-related apoptosis inducing ligand (TRAIL). After isolation, characterization and transfection, the authors show proper expression of TRAIL, migration of MSCs toward pancreatic cancer cells and cytotoxicity in pancreatic cancer cells.

TRAIL-engineered pancreas-derived mesenchymal stem cells: characterization and cytotoxic effects on pancreatic cancer cells
MR Moniri, X-Y Sun, J Rayat, D Dai, Z Ao, Z He, CB Verchere, L-J Dai and GL Warnock
Cancer Gene Ther. 2012 Sep;19(9):652-8. doi: 10.1038/cgt.2012.46. Epub 2012 Jul 6.

What is the significance of TRAIL?

Induction of apoptosis is an effective means by which many chemotherapeutic agents kill cancer cells. There are two pathways, intrinsic and extrinsic, by which apoptosis can be induced. The intrinsic pathway is generated via intracellular signals from the mitochondria in response to chemotherapy, radiation or other intracellular changes (e.g. DNA damage). Extrinsic apoptosis induction is activated via cell surface death receptors (DRs) that belong to the TNF superfamily which also includes TRAIL receptors. TRAIL also possesses an ideal characteristic of inducing apoptosis in transformed or tumor cells but not normal cells.

To examine TRAIL induced apoptosis, MSCs and pancreatic cancer cells used in this study were first characterized. Upon successful isolation of pancreas-derived MSCs, these cells were differentiated into adipogenic and osteogenic cells and characterized through biomarker expression (e.g. CD44, CD73, CD95, and CD105) prior to transfection with TRAIL-bearing plasmids. In addition, human pancreatic cells from ATCC (BXPC3, ASPC1 and PANC1) and transformed human islet cells (HP62 and TRM6) were characterized through flow cytometry of TRAIL receptors – DR4, DR5, Dcr1 and DcR2.

Cancer cell toxicity from TRAIL expressing MSCs

Viability and migration experiments of cancer cells were performed via an impedance-based real-time cell analyzer (RTCA). Growth profiles for each cancer cell were generated to find the optimal time point and cell density at which to test cytotoxicity to avoid false-positive or false-negative results. After titrating different cell densities, a 20,000 cells/well and 20h time point was chosen. With this reference established, co-cultures of MSCs and pancreatic cancer cells were seeded within the RTCA system to show dynamic migration of MSCs toward each pancreatic cell line. Migration effects were further enhanced in the presence of conditioned media.

Further experiments go on to show cytotoxicity on pancreatic cancers cells from MSCs expressing nsTRAIL (non-secreting) and stTRAIL (secreting). In particular, ASPC1, TRM6 and HP62 cell types displayed significant reduction in viability from MSCs in the presence of conditioned media. MSCs transfected with secreting TRAIL were also more effective in reducing viability in pancreatic cancer cells than were cells transfected with non-secreting TRAIL.

Implications

Pancreatic cancer continues to have a very poor prognosis and treatments are elusive due to the lack of tumor specific therapies; however, the tumor-homing characteristic of MSCs provides an attractive vehicle to deliver therapeutics. The current study shows this proof of concept through expression of the anti-cancer agent TRAIL from MSCs transfected using TransIT®-2020.

View more information on transfection of stem cells.

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