Targeting Mutational Drivers in IPMN Using Lipid Nanoparticle Delivery Systems
Abstract
Background: Unlike other pancreatic ductal adenocarcinoma (PDAC) precursors, ~90% of intraductal papillary mucinous neoplasms (IPMN) are diagnosed before cancer is present, providing a unique window of opportunity for therapeutic intervention before PDAC forms. Overall, 96% of IPMNs harbor KRAS (~80%) and/or GNAS (~66%) driver mutations. Although SMAD4 inactivation occurs in only 13% of IPMN, loss of expression is associated with progression to advanced stage IPMN in humans and mouse models. Oncogenic drivers such as KRAS and GNAS have proven difficult to target and there are currently no therapies designed to restore tumor suppressor function. Our mutation-specific siRNAs against KRASG12D and GNASR201C target oncogenic drivers in IPMN while minimizing any potential off-target effects. Our DLin-MC3-DMA-pegylated lipid nanoparticles (LNPs) are the most clinically advanced delivery system and the primary receptor for LNPS, low-density lipoprotein receptor, is overexpressed in mouse and human IPMN. We propose to utilize LNP-RNA targeting in IPMN models to resolve IPMN pre-malignant lesions before they progress to cancer, preventing unnecessary surgery. FY21 PCARP Focus Areas: (1) Understanding precursors, origins, and early progression of pancreatic cancer, (2) Understanding the events that promote pancreatic cancer metastasis, (3) New drug development targeted toward cancer sensitivity and resistance mechanisms. Hypothesis/Objective: Mutation-specific LNP-siRNAs targeting KRAS and/or GNAS or LNP-mRNAs that restore SMAD4 expression will block the progression of IPMN to PDAC. Our objectives are to determine the efficacy and safety of LNP-RNA therapies in multiple preclinical models of IPMN. Specific Aims: Aim 1. To delineate the interrelated mechanisms by which KRASG12D, GNASR201C, and SMAD4 drive IPMN progression. Aim 2. To investigate the uptake, therapeutic efficacy, and safety of LNP-siRNAs targeting KrasG12D and/or GnasR201C in a mouse model of IPMN. Aim 3. To determine the dependence of IPMN maintenance and progression on KrasG12D and/or Smad4 loss. Study Design: Aim 1: IPMN-derived PDAC cell lines will be treated with mutation-specific LNP- siRNA, LNP-mRNA or controls alone or in combination and will then undergo RNA sequencing to identify altered downstream signaling pathways. Aim 2: We will evaluate and optimize LNP-siRNA delivery to premalignant IPMN in Kras;Gnas mice, an autochthonous model of IPMN. Aim3: We will use IPMN-derived PDAC cell lines to determine the effects of LNP-mRNA for SMAD4 on restoring protein expression, down-stream target expression, and on cell survival and proliferation. We will use a novel autochthonous mouse model of IPMN, Kras;Smad4, to test the use of combined LNP-siRNA and LNP-mRNA as a therapeutic strategy to block the progression of IPMN to PDAC. Impact: Due to overtreatment bias, almost half of IPMN patients without malignancy undergo surgery and suffer from surgery-associated complications. Our proposal addresses this major clinical need by utilizing U.S. Food and Drug Administration-approved LNPs to deliver mutation-specific siRNA/mRNA into the cyst via systemic administration or widely used clinical ultrasound guided imaging to eliminate pre-malignant lesions without unnecessary surgery. Additionally, we will uncover alterations in signaling pathways, which will increase our understanding of PDAC and may identify new, druggable targets. Our vision is to translate this work into clinical trials utilizing this technology to target cancer-specific and/or patient-specific genes in pancreatic cancer. Innovation: Our LNP-RNA technology for combination targeting of mutant-driver genes and to restore the expression of tumor suppressor genes is a novel strategy for the treatment of pancreatic cancer. Results obtained from our proposed studies will help fill in the gaps in understanding on how KRAS, GNAS, and SMAD4 mutations independently and synergis
Document Details
- Document Type
- DoD Grant Award
- Publication Date
- Dec 28, 2022
- Source ID
- W81XWH2211122
Entities
People
- Nidhi Jyotsana
Organizations
- United States Army
- Vanderbilt University