SBIR-STTR Award

c-MYC has been recognized for three decades as a key oncogenic driver since the discovery of retroviral oncogenes, their normal cellular counterparts, and immediate early response genes that are activated in response to mitogenic factors. A large fraction of all human cancers have also been shown to contain c- MYC gene amplifications, including 100% of some hematologic tumors and up to 70% of all solid tumors. Genetically engineered tumors of multiple types have confirmed the profound roll of c-MYC in tumor initiation and progression, and down regulation of c-MYC expression has shown significant therapeutic effects in many GEMM tumor types. Unlike most successfully drugged disease-causing proteins, c-MYC does not participate directly in a cellular signaling cascade as a receptor, kinase, or other signal transduction element and the protein has no known enzymatic activity or other function suitable for small molecule binding. Due to the inherent difficulty in targeting protein-protein and protein-DNA binding interactions, c-MYC has remained undruggable by conventional approaches. RNAi is a modality that can overcome the challenges of drugging the c-MYC protein directly because RNAi targets and destroys messenger RNA in a sequence-specific manner, thus reducing target protein expression as a consequence of mRNA elimination. RNA duplexes dissolved in infusion media and systemically administered as "free" duplexes are ineffective as cancer therapeutic agents for three reasons: 1) rapid clearance from blood by renal filtration, 2) rapid degradation by ubiquitous nucleases in blood, and 3) lack of accumulation in tumor tissues. One approach to overcome all three of these therapeutic limitations is to develop clinically acceptable nanoparticle formulations to efficiently deliver RN to tumors in vivo. Lipid-based nanoparticles are the most well established clinical stage formulations due to their similarities to approved liposomal drug products. By focusing our research and development on tumor-bearing animals, we have developed both a novel lipid composition and manufacturing process that dramatically improves both efficacy and tolerability of LNPs for RNA delivery, while minimizing inflammatory effects. Thus, our c-MYC DsiRNA formulated in LNPs that mediate intratumoral delivery has the potential to yield a therapeutic response in up to 70% of all cancer patients and with an acceptable therapeutic index. In this application we seek to expand the therapeutic index of our experimental drug further, enabling it to be used in patients highly compromised by their disease state, prior therapies, or in the case of hepatocellular carcinoma patients, by underlying chronic liver disease. Project Description Page 6.

Public Health Relevance Statement:


Public Health Relevance:
We will optimize both the approved pharmaceutical ingredient in a DsiRNA-based therapeutic targeting the cMYC oncogene, and the lipid nanoparticle delivery system itself by using a new functional excipient.

Project Terms:
Amides; Animals; antitumor agent; base; Binding (Molecular Function); Biocompatible; Biological Assay; Blood; Cancer Model; Cancer Patient; Chemicals; chronic liver disease; Clinical; Clinical Trials; Development; Disease; DNA Binding; Down-Regulation; Drug Delivery Systems; Drug Formulations; Elements; ERG gene; Ethers; Excipients; Failure (biologic function); Filtration; Gene Dosage; Genes; Genetic Engineering; Genetic Models for Cancer; Genetic Transcription; Hepatitis B Virus; Hepatitis C; Human; improved; In Vitro; in vivo; Inflammatory; Infusion procedures; Interleukin-2; intravenous injection; Kidney; Knowledge; Korea; Life; Link; Lipids; Liposomes; Literature; Liver; liver function; Maintenance; Malignant Neoplasms; manufacturing process; Marketing; Mediating; Messenger RNA; metaplastic cell transformation; Micelles; Modality; Modeling; Modification; Mortality Vital Statistics; Mus; Mutation; MYC Family Protein; MYC Gene Amplification; nanoparticle; neoplastic cell; nonhuman primate; novel; nuclease; Oncogenes; Oncogenic; oncology; Patients; Pharmaceutical Preparations; Pharmacologic Substance; Phase; phase 1 study; Phase I Clinical Trials; Phosphotransferases; Preparation; Primary carcinoma of the liver cells; Prior Therapy; Process; protein expression; protein function; Proteins; public health relevance; Qualifying; receptor; Refractory; research and development; response; RNA; RNA Interference; Role; Safety; Signal Transduction; Site; small molecule; Solid Neoplasm; Staging; System; Taiwan; Technology; Therapeutic; Therapeutic Agents; Therapeutic Effect; Therapeutic Index; therapeutic target; TLR4 gene; Toxicology; tumor; Tumor Expansion; tumor initiation; tumor progression; Tumor Tissue; tumor xenograft

Hepatocellular carcinoma (HCC) is the third most common cause of cancer-related deaths worldwide, with the incidence increasing significantly in the West. The prognosis for HCC is poor, and the limited success of the standard of care drug sorafenib along with recent Phase III failures of other treatments underscores the need for novel HCC therapeutics. Intrahepatic cholangiocarcinoma (IHCC), the second most frequent primary liver tumor, is also associated with poor prognosis. Colorectal cancer (CRC) is the fourth most common cause of cancer deaths throughout the world. Despite the recent advances in treatment strategies, post-operative prognosis of CRC patients with liver, lymph node, or distant organ metastasis remains poor, underscoring the need for novel therapeutics. The Wnt signaling pathway is commonly activated in all three of these diseases, and this is thought to be a causal event in their tumorigenesis. In this application we continue the development of a lipid nanoparticle (LNP) platform, using an RNAi- based payload that targets a key component of the Wnt signaling pathway, beta-catenin (CTNNB1). Beta- catenin, which is well-validated as an oncology target with high clinical mutation occurrence and experimental evidence, is considered undruggable via conventional approaches. However, by enabling pharmacological intervention at the mRNA level as opposed to the protein level, our RNAi approach inhibits synthesis of beta- catenin protein and causes tumor growth inhibition in preclinical models. Building upon the successful development of our LNP-encapsulated Dicer Substrate siRNA (siRNA) targeting the MYC oncogene, we sought to improve LNP formulations and demonstrate effectiveness with a second payload, delivering CTNNB1 to tumors. Our in-house experience with developing and cGMP manufacturing LNP-formulated therapeutics, as well as the similarities to approved liposomal oncology products, led us to generate and characterize LNPs with unique chemical compositions carrying novel CTNNB1 DsiRNA payloads. Here we demonstrate dramatic improvement in efficacy and other properties of tumor-centric LNPs. Through this work, a formulated LNP-CTNNB1 DsiRNA candidate that is suitable for preclinical development and IND submission has been identified. In this application we seek to perform IND enabling studies, process development, and GMP manufacture of this candidate, as well as perform translational research to guide the clinical research strategy. The ultimate goal is to develop a Lipid Nanoparticle formulation that can be used to deliver a broad range of siRNA payloads, with a tolerability profile to enable treatment of patients with a spectrum of disease states and prior therapeutic histories.

Public Health Relevance Statement:


Public Health Relevance:
We will continue the preclinical development of lipid nanoparticle delivery platform, demonstrating efficacy with a DsiRNA that targets the Wnt signaling pathway in tumors.

NIH Spending Category:
Bioengineering; Biotechnology; Cancer; Colo-Rectal Cancer; Digestive Diseases; Genetics; Liver Cancer; Liver Disease; Nanotechnology; Orphan Drug; Rare Diseases

Project Terms:
base; BAY 54-9085; beta catenin; Biological Assay; Biopsy; Cancer Etiology; Cellular biology; Cessation of life; Chemicals; Clinic; Clinical; Clinical Research; colon cancer patients; Colorectal Cancer; Combined Modality Therapy; CTNNB1 gene; Cyclic GMP; Development; Disease; Distant; Dose; Dose-Limiting; Drug Delivery Systems; Effectiveness; Encapsulated; Epigenetic Process; Event; experience; Failure; Formulation; Freezing; Genetic Models; Goals; Growth; Housing; Immunomodulators; improved; Incidence; Intervention; Intrahepatic Cholangiocarcinoma; Lipids; Liposomes; Liver; Liver neoplasms; lymph nodes; Measurement; meetings; Messenger RNA; Methods; Modeling; Molecular; molecular subtypes; Mutation; nanoparticle; Neoplasm Metastasis; novel; novel therapeutics; Oncogenes; oncology; Organ; outcome forecast; Pathway interactions; Patients; Pharmaceutical Preparations; Phase; Postoperative Period; pre-clinical; Pre-Clinical Model; Primary carcinoma of the liver cells; Process; Property; Proteins; public health relevance; Recording of previous events; Regimen; Research; research and development; RNA Interference; scale up; Signal Pathway; Small Business Innovation Research Grant; Small Interfering RNA; Solid Neoplasm; stability testing; Staging; standard of care; success; Surveys; Technology; Therapeutic; therapeutic target; Tissues; tool; Toxic effect; Toxicology; Translational Research; treatment strategy; tumor; tumor growth; tumor microenvironment; tumorigenesis; Work