PBAE-PEG/Lipid Nanoparticle Delivery of RNA for the Creation of Genetically Engineered Lung Cancer Mouse Models

Abstract

We have recently developed polymer/lipid nanoparticles PBAE-PEG/4A3-SC8/DOPE/Cholesterol/DOTAP (hereafter, PBAE-PEG/LNP) that can deliver mRNA into lung cells. Here, using PBAE-PEG/LNP, we delivered Cre mRNA and/or sgRNAs into Kras^LSL-G12D/+ and/or Cas9 mice to develop genetically engineered lung cancer mouse models. PBAE-PEG/LNP delivery of Cre mRNA into Kras^LSL-G12D/+;Cas9 mice by intratracheal (IT) injection produced autochthonous lung tumors while intravenous injection resulted in lung tumors as well as bronchus-associated lymphoid tissue (BALT). PBAE-PEG/LNP delivery of Cre mRNA along with sgRNA targeting the lung lineage transcription factor Nkx2-1 (sgNkx2-1) into Kras^LSL-G12D/+;Cas9 mice by IT injection produced autochthonous invasive mucinous adenocarcinoma of the lung (IMA) that lacks NKX2-1 while expressing the gastrointestinal transcription factor HNF4A. PBAE-PEG/LNP delivery of sgRNAs targeting Eml4 (sgEml4) and Alk (sgAlk) into Cas9 mice by IT injection produced autochthonous lung tumors carrying the driver oncogene Eml4-Alk. This approach using PBAE-PEG/LNP to deliver RNA will allow for agile development of lung cancer mouse models.

Keywords: ALK; GEMM; KRAS; LNP; NKX2-1; lung cancer.