Pilot Project 3 - Lung

This is a proposal in which a medical biochemist from FAMU (Dr. Lamango), a biomedical engineer from UF (Dr. Huang), and a molecular geneticist from USC (Dr. Offringa) combine their innovative resources to tackle lung cancer in Blacks. Lung adenocarcinoma (LUAD) is the most common lung cancer subtype in all ethnic/racial groups, including Blacks. Cell line models should be available for all racial/ethnic groups, since genetic background could affect driver gene function and response to therapies. However, in vitro systems to study LUAD development and test therapeutics in Black Americans are sorely lacking. We hypothesize that due to genetic differences, LUAD in Blacks will have a unique repertoire of cancer driver genes and will respond to targeted therapies distinctly from White LUAD. There are no immortalized alveolar epithelial cells from Blacks and only 1 Black LUAD cell line, yet such cells are indispensable to study LUAD development and the therapeutic disparity in lung cancer treatment. This project aims to begin to fill this critical gap through three Specific Aims. Aim 1 is to create a novel collection of cell lines specifically tailored to the study of lung cancer in Blacks. We will develop immortalized alveolar epithelium cell lines from unidentified Black subjects as well as isogenic LUAD cell lines with various driver mutations from Black subjects. Aim 2 is to use 3 dimensional (3D) printing technology to engineer an alveolar organoid for rapid drug screening and testing. We will establish a device with a simulated air sac, surrounded by simulated blood vessels. This will allow manipulation of lung cells through the airway inlet (e.g. using the Cre-lox system and Adenovirus-Cre delivery through the air interface) and screening of drugs through the blood inlet. Aim 3 is to test polyisoprenylated cysteinyl amide inhibitors (PCAIs), agents shown in the Lamango lab to disrupt the RAS-mediated signaling (a pathway frequently altered in LUAD) in ethnically/racially diverse cell lines. Work will be initiated using existing cell line H23 and for comparison other LUAD cell lines from non-Black individuals using in vitro cell culture studies. As new cell lines are established from Aim 1, these will be incorporated into the study. As the organoids arise from Aim 2, the drugs will also be tested in that model system. Each of the three specific Aims can generate important novel data/reagents on its own, including new alveolar cell lines and isogenic lung cancer cell lines from Black Americans, a unique new engineered 3D-printed alveolar organoid system, and testing of PCAIs on lung cancer cells with and without KRAS mutations. In addition, in synergy the new cell lines from Aim 1 can be used in the newly developed alveolar organoid system (Aim 2) and will allow testing of PCAIs on an expanded cell line collection and organoids (Aim 3). These independent yet synergistic aims will provide preliminary data that will lay the foundation for a strong R01 application by this team, such as for PAR 18-654, Basic Research in Cancer Health Disparities, and similar future RFAs.