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Enhanced anti-tumor efficacy with multi-transgene armed mesenchymal stem cells for treating peritoneal carcinomatosis
Yoon Khei Ho1,2,3,4, Jun Yung Woo5,6, Kin Man Loke5,6
1Department of Biochemistry, National University of Singapore, Singapore, 117596, Singapore. bchhyk@nus.edu.sg.
Background:
Mesenchymal stem cells (MSCs) have garnered significant interest for their tumor-tropic property, making them potential therapeutic delivery vehicles for cancer treatment. We have previously shown the significant anti-tumour activity in mice preclinical models and companion animals with naturally occurring cancers using non-virally engineered MSCs with a therapeutic transgene encoding cytosine deaminase and uracil phosphoribosyl transferase (CDUPRT) and green fluorescent protein (GFP). Clinical studies have shown improved response rate with combinatorial treatment of 5-fluorouracil and Interferon-beta (IFNb) in peritoneal carcinomatosis (PC). However, high systemic toxicities have limited the clinical use of such a regime.
Methods:
In this study, we evaluated the feasibility of intraperitoneal administration of non-virally engineered MSCs to co-deliver CDUPRT/5-Flucytosine prodrug system and IFNb to potentially enhance the cGAS-STING signalling axis. Here, MSCs were engineered to express CDUPRT or CDUPRT-IFNb. Expression of CDUPRT and IFNb was confirmed by flow cytometry and ELISA, respectively. The anti-cancer efficacy of the engineered MSCs was evaluated in both in vitro and in vivo model. ES2, HT-29 and Colo-205 were cocultured with engineered MSCs at various ratio. The cell viability with or without 5-flucytosine was measured with MTS assay. To further compare the anti-cancer efficacy of the engineered MSCs, peritoneal carcinomatosis mouse model was established by intraperitoneal injection of luciferase expressing ES2 stable cells. The tumour burden was measured through bioluminescence tracking.
Results:
Firstly, there was no changes in phenotypes of MSCs despite high expression of the transgene encoding CDUPRT and IFNb (CDUPRT-IFNb). Transwell migration assays and in-vivo tracking suggested the co-expression of multiple transgenes did not impact migratory capability of the MSCs. The superiority of CDUPRT-IFNb over CDUPRT expressing MSCs was demonstrated in ES2, HT-29 and Colo-205 in-vitro. Similar observations were observed in an intraperitoneal ES2 ovarian cancer xenograft model. The growth of tumor mass was inhibited by ~ 90% and 46% in the mice treated with MSCs expressing CDUPRT-IFNb or CDUPRT, respectively.
Conclusions:
Taken together, these results established the effectiveness of MSCs co-expressing CDUPRT and IFNb in controlling and targeting PC growth. This study lay the foundation for the development of clinical trial using multigene-armed MSCs for PC.
Insights
Engineered mesenchymal stem cells (MSCs) co-delivering cytosine deaminase and uracil phosphoribosyl transferase (CDUPRT) with Interferon-beta (IFNb) show potent anti-cancer activity. This combination therapy effectively targets peritoneal carcinomatosis growth with reduced toxicity.
Area of Science:
- Oncology
- Stem Cell Therapy
- Gene Therapy
Background:
- Mesenchymal stem cells (MSCs) are explored as cancer delivery vehicles due to their tumor-homing properties.
- Previous studies demonstrated anti-tumor effects of MSCs engineered with cytosine deaminase and uracil phosphoribosyl transferase (CDUPRT).
- Combinatorial therapy with 5-fluorouracil and Interferon-beta (IFNb) shows promise for peritoneal carcinomatosis (PC) but is limited by systemic toxicities.
Purpose of the Study:
- To evaluate the feasibility of using intraperitoneally administered, non-virally engineered MSCs for co-delivery of a CDUPRT/5-Flucytosine prodrug system and IFNb.
- To investigate the potential enhancement of the cGAS-STING signaling axis through this combined MSC-based therapy.
- To assess the anti-cancer efficacy of MSCs engineered to express CDUPRT or CDUPRT-IFNb in vitro and in vivo.
Main Methods:
- MSCs were engineered to express either CDUPRT or a fusion protein CDUPRT-IFNb.
- Expression of transgenes was confirmed using flow cytometry and ELISA.
- In vitro efficacy was tested using co-culture assays with ovarian cancer cells (ES2, HT-29, Colo-205) and MTS assay for cell viability.
- In vivo efficacy was evaluated in a peritoneal carcinomatosis mouse model using luciferase-expressing ES2 cells, with tumor burden monitored by bioluminescence.
Main Results:
- Engineered MSCs maintained their phenotype and migratory capabilities despite high transgene expression.
- MSCs co-expressing CDUPRT-IFNb demonstrated superior anti-cancer efficacy compared to CDUPRT-expressing MSCs in vitro.
- In vivo, MSCs expressing CDUPRT-IFNb inhibited tumor mass growth by approximately 90%, significantly outperforming MSCs expressing CDUPRT (46% inhibition).
Conclusions:
- MSCs engineered to co-express CDUPRT and IFNb are effective in controlling and targeting peritoneal carcinomatosis growth.
- This multigene-armed MSC approach offers a promising strategy for treating PC.
- The findings provide a foundation for developing clinical trials utilizing genetically modified MSCs for peritoneal carcinomatosis treatment.
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