The miR15a/16 Axis and Precision Gene Intervention to Provide a Novel Therapy for Lung Cancer
Abstract
A recent advance in cell biology is the discovery of microRNA: small, non-coding nucleotide strands that have the potential to negatively regulate the expression of potentially hundreds of genes by the process of RNA interference. These miRNAs have been demonstrated to play a role in numerous cellular processes related to cancer, including cell differentiation, proliferation, and migration. The loss of expression of two specific and cotranscribed species, miR15a and miR16, has been identified in numerous cancers, including those of the lung, although their precise role in the initiation and progression of cancer has not been fully elucidated. miRlSa/16 are responsible for the regulation of several well-defined oncogenic pathways, The goal of this proposal is to develop a highly specific therapy capable of fighting lung cancer, with the central hypothesis that loss of miRlSa/16 is a triggering event in the progression of healthy cells to a cancerous state, and that rescue of miRlSa/16 expression will effectively halt growth and metastasis in lung cancer cells. In order to do so, we propose two specific aims. Specific Aim 1 (SAl) will demonstrate the effect of targeted CRISPR/Cas9 transfection of miRlSa/16 specific to SCLC and NSCLC in vitro. This aim will use the CRISPR/Cas9 system to insert a functional miRlSa/16 sequence into the cancer cell genome, with the goal of fundamentally altering the nature of these malignant cells. To ensure cancer cell-specific targeting, vectors coding for the Cas9 enzyme and miRlSa/sequence will be delivered using a lentiviral vector that has been conjugated to an antibody for MAGE-3, a cell surface protein that has been demonstrated to be uniquely expressed in both SCLC (H446) and NSCLC (A549), in order to promote fusion of the lentivirus with the cell membrane and subsequently insert a miRlSa/16 sequence into the cancer cell genome using a CRISPR/Cas9 genetic editing technique, restoring production of these regulatory microRNA.
Document Details
- Document Type
- Technical Report
- Publication Date
- Oct 01, 2023
- Accession Number
- AD1225472
Entities
People
- James D. Fluckey
- Peter Nghiem
Organizations
- Texas A&M University