Enhanced gene transfection of macrophages by photochemical internalization: Potential for gene-directed enzyme prodrug therapy of gliomas

Gabrielle Romena; Lina Nguyen; Kristian Berg; Steen J Madsen; Henry Hirschberg

doi:10.1016/j.pdpdt.2020.102098

Enhanced gene transfection of macrophages by photochemical internalization: Potential for gene-directed enzyme prodrug therapy of gliomas

Photodiagnosis Photodyn Ther. 2021 Mar:33:102098. doi: 10.1016/j.pdpdt.2020.102098. Epub 2020 Nov 11.

Authors

Gabrielle Romena¹, Lina Nguyen², Kristian Berg³, Steen J Madsen⁴, Henry Hirschberg⁵

Affiliations

¹ Beckman Laser Institute and Medical Clinic, University of California, Irvine 1002 Health Sciences Rd, Irvine, CA, 92617, USA. Electronic address: gromena@uci.edu.
² Beckman Laser Institute and Medical Clinic, University of California, Irvine 1002 Health Sciences Rd, Irvine, CA, 92617, USA.
³ Dept. of Radiation Biology, The Norwegian Radium Hospital, Oslo University Hospital, Montebello, N-0310, Oslo, Norway.
⁴ Dept. of Health Physics and Diagnostic Sciences, University of Nevada, Las Vegas, NV 89154, USA.
⁵ Beckman Laser Institute and Medical Clinic, University of California, Irvine 1002 Health Sciences Rd, Irvine, CA, 92617, USA; Dept. of Health Physics and Diagnostic Sciences, University of Nevada, Las Vegas, NV 89154, USA.

PMID: 33188941
DOI: 10.1016/j.pdpdt.2020.102098

Abstract

Background: Drawn by tumor synthesis of chemo-attractive factors, macrophages are frequently found in and around glioblastomas and play an important role both in augmenting as well as inhibiting tumor growth. Patient-derived macrophages have the potential, therefore, to act as targeted delivery vectors for a variety of anti-cancer treatments. Among these is ex vivo gene transfection and re-injection back into the patient of macrophages to target residual tumors. In this study, photochemical internalization (PCI) is investigated as a technique for the non-viral transfection of the cytosine deaminase (CD) prodrug activating gene into macrophages. The CD gene encodes an enzyme that converts the nontoxic antifungal agent, 5-fluorocytosine (5-FC), into 5-fluorouracil (5-FU) - a potent chemotherapeutic agent.

Materials: PCI (photosensitizer + light treatment) mediated CD gene transfection of rat alveolar Ma cells was carried out in vitro. CD gene transfected NR8383 macrophages were co-cultured with F98 rat glioma cells in the presence or absence of 5-FC. Cell viability was assayed using the MTS colorimetric assay.

Results: Compared to the glioma cells, NR8383 demonstrated enhanced resistance to the toxic effects of 5-FU. PCI greatly increased the transfection efficiency of the CD gene in NR8383 cells. The viability of F98 cells was significantly inhibited by coculture with CD transfected NR8383 macrophages and 5-FC.

Conclusion: Although gene insertion into macrophages has proven difficult, the results presented here show that non-viral transfection of the CD gene into these immune cells can be enhanced via PCI. CD transfected NR8383 cells could efficiently convert 5-FC to 5-FU and export the drug, producing a pronounced bystander toxic effect on adjacent non-transfected glioma cells. Compared to single treatment, repetitive PCI-induced transfection was more efficient at low CD plasmid concentration.

Keywords: 5-fluorouracil; E. coli cytosine deaminase; Gene-directed enzyme prodrug therapy; Gliomas; Photochemical internalization.

MeSH terms

Animals
Cell Line, Tumor
Cytosine Deaminase / genetics
Fluorouracil
Glioma* / drug therapy
Glioma* / genetics
Humans
Macrophages
Photochemotherapy* / methods
Photosensitizing Agents / pharmacology
Photosensitizing Agents / therapeutic use
Prodrugs* / therapeutic use
Rats
Transfection

Substances

Photosensitizing Agents
Prodrugs
Cytosine Deaminase
Fluorouracil