Engineering nucleic acid constructs for lymphoma therapy

Abstract

Serine is an essential metabolite in cancer cells. In addition to being imported from the extracellular environment, serine can be synthesized through de novo serine synthesis (DNSS), a pathway that branches off glycolysis. Interestingly, unlike normal cancer cells, Natural killer/T cell lymphoma (NKTL) does not express phosphoglycerate dehydrogenase (PHGDH), the rate-limiting enzyme of DNSS. However, NKTL patients who experience prolonged survival have higher tumoral PHGDH expression levels, suggesting PHGDH may offer an advantage to NKTL patients by affecting the growth of NKTL cells. Previous experiments in vitro introducing PHGDH into NKS1 did not affect their growth and viability. To aid ongoing laboratory efforts in understanding the role of PHGDH in NKTL in vivo, we developed a workflow for producing transfection-competent PHGDH mRNA-lipid complexes. The synthesis, lipid encapsulation, and transfection of PHGDH mRNA were achieved in NKYS. However, transfection was unsuccessful in NKS1 and activated primary T and NK cells. The transfection of PHGDH mRNA-lipid complexes into NKTL and primary T and NK cells will allow us to study their impact on cell functionality, including cytokine production. Our goal is to utilize these complexes for intratumoral injections in vivo to uncover the effects of PHGDH on tumor growth.