A highly efficient non-viral process for programming mesenchymal stem cells for gene directed enzyme prodrug cancer therapy

Abstract

Mesenchymal stem cells (MSCs) driven gene-directed enzyme prodrug therapy has emerged as a potential strategy for cancer treatment. The tumour-nesting properties of MSCs enable these vehicles to target tumours and metastases with effective therapies. A crucial step in engineering MSCs is the delivery of genetic material with low toxicity and high efficiency. Due to the low efficiency of current transfection methods, viral vectors are used widely to modify MSCs in preclinical and clinical studies. We show, for the first time, the high transfection efficiency (> 80%) of human adipose tissue derived-MSCs (AT-MSCs) using a cost-effective and off-the-shelf Polyethylenimine, in the presence of histone deacetylase 6 inhibitor and fusogenic lipids. Notably, the phenotypes of MSCs remained unchanged post-modification. AT-MSCs engineered with a fused transgene, yeast cytosine deaminase::uracil phosphoribosyltransferase (CDy::UPRT) displayed potent cytotoxic effects against breast, glioma, gastric cancer cells in vitro. The efficiency of eliminating gastric cell lines were effective even when using 7-day post-transfected AT-MSCs, indicative of the sustained expression and function of the therapeutic gene. In addition, significant inhibition of temozolomide resistant glioma tumour growth in vivo was observed with a single dose of therapeutic MSC. This study demonstrated an efficient nonviral modification process for MSC-based prodrug therapy.

Modified mesenchymal stem cells in cancer therapy: A smart weapon requiring upgrades for wider clinical applications

“…Polyethylenimine based polymers were used to transiently engineer MSC with HSV-TK, together with TRAIL: These modified cells were effective in vitro and in vivo against glioma through increased apoptosis and reduced angiogenesis[ 178 ]. MSC expressing CDy::UPRT by the same transfection method significantly inhibited in vivo temozolomide resistant glioma tumors[ 179 ] as well as 5-fluorouracil resistant colorectal adenocarcinoma cells[ 180 ].…”

Current Status of Mesenchymal Stem/Stromal Cells for Treatment of Neurological Diseases

“…Several virus-based approaches have been used for genetic engineering of MSCs, including retrovirus, lentivirus, and adenovirus, each of which has advantages and limitations. Recently, an efficient non-viral process using a cationic polymer for MSC programming has been described (Ho et al, 2020 ). Other approaches, such as plasmid transfection, can also be used to modify MSCs, but viral vectors are still the most widely used tool for genetic engineering of MSCs in preclinical trials, as in the majority of studies described in this section.…”

Mesenchymal stem cell‐glioblastoma interactions mediated via kinin receptors unveiled by cytometry

“…Thus, primed MSCs or MSCs loaded with anti‐tumor agents can be considered feasible for GBM therapy. For example, cytosine deaminase fused with uracil phosphoribosyltransferase (CDy::UPRT) expressing MSCs have shown anti‐tumor effects in several cancer types [6–8]. According to Altaner and colleagues, the infusion of CDy::UPRT‐transfected MSCs after GBM resection may present a valid strategy to prevent tumor relapse [6].…”

“…According to Altaner and colleagues, the infusion of CDy::UPRT‐transfected MSCs after GBM resection may present a valid strategy to prevent tumor relapse [6]. Suppression of U251 tumor growth has been demonstrated by injecting CDy::UPRT‐transfected MSCs with 5‐fluorocytosine (5‐FC), a widely used antifungal prodrug, directly into the subcutaneous tumor [7, 8]. MSCs can also be engineered to secrete higher amounts of exosomes containing antitumor miRNAs and proteins that inhibit tumor growth after systemic administration [76].…”

“…MSCs are endogenous, immunomodulatory and mobile tumor‐tropic cells that can be harvested from adipose tissue, oral cavity, umbilical cord and various other tissues with the least discomfort for the patient. They are easily propagated and engineered in vitro and have shown promising results as anti‐GBM agents [6–8]. Although, not only beneficial traits, but also tumorigenic properties were reported regarding MSCs in oncology studies [9–11].…”