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 <i>Cre</i> mRNA and/or sgRNAs into <i>Kras</i>^{<i>LSL-G12D/+</i>} and/or <i>Cas9</i> mice to develop genetically engineered lung cancer mouse models. PBAE-PEG/LNP delivery of <i>Cre</i> mRNA into <i>Kras</i>^{<i>LSL-G12D/+</i>}<i>;Cas9</i> 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 <i>Cre</i> mRNA along with sgRNA targeting the lung lineage transcription factor <i>Nkx2-1</i> (sgNkx2-1) into <i>Kras</i>^{<i>LSL-G12D/+</i>}<i>;Cas9</i> 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 <i>Eml4</i> (sgEml4) and <i>Alk</i> (sgAlk) into <i>Cas9</i> mice by IT injection produced autochthonous lung tumors carrying the driver oncogene <i>Eml4-Alk</i>. This approach using PBAE-PEG/LNP to deliver RNA will allow for agile development of lung cancer mouse models.