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Principal Investigator: William Raymond Lagor
Organization: BAYLOR COLLEGE OF MEDICINE
Fiscal Year: 2024
Award: $547,307
Funding agency: National Heart Lung and Blood Institute
This is a competing renewal R01 application that seeks to develop new gene editing approaches to treat Familial Hypercholesterolemia (FH). FH is an autosomal dominant disease most often caused by loss-of-function mutations in the low density lipoprotein receptor (LDLR), the protein responsible for uptake of ApoB-containing lipoprotein particles (such as LDL) by the liver. Loss of both LDLR alleles in homozygous FH (HoFH) causes excessively high plasma cholesterol (~1000 mg/dl), xanthomas, atherosclerosis, and death in the first few decades of life if untreated. Currently, the only curative treatment for HoFH is a liver transplant, making this disease an important priority for gene therapy. Achieving permanent correction of HoFH through gene therapy will require modifying the patient’s own DNA, so that cells expressing LDLR will persist in the liver. In this application we will further expand on our previous work, by developing strategies to integrate a full length LDLR transgene into genomic safe harbor sites. The goal is to ensure that the therapy will be generalizable to HoFH patients, and not specific to a particular mutation or region of the protein. Secondly, we will use FH mouse models to test a new strategy for liver-directed gene therapy through selection expansion of gene-corrected hepatocytes. This approach capitalizes on the regenerative capacity of the liver, and uses endogenous essential gene as a selectable marker. Over time, inhibition of the essential gene can be used to selectively expand gene-targeted hepatocytes, ensuring that liver-wide correction can be achieved. We propose two specific Aims: 1) Determine if targeted integration at the Ldlr and Apoa1 loci can correct FH, and 2) Develop a novel system for selective expansion of gene- corrected hepatocytes. Completion of this work will produce a new gene therapy solutions for HoFH, as well as a universal system for genetic correction of other liver disorders through in vivo selection.
Terms: <Acids><Alleles><Allelomorphs><Apheresis><Apo-B><Apo-E><ApoB><ApoE><ApoE protein><Apolipoprotein E><Apolipoproteins B><Apoptosis><Apoptosis Pathway><Atherosclerosis><Atherosclerotic Cardiovascular Disease><Blood><Blood Component Removal><Blood Plasma><Blood Reticuloendothelial System><CRISPR><CRISPR approach><CRISPR based approach><CRISPR method><CRISPR methodology><CRISPR technique><CRISPR technology><CRISPR tools><CRISPR-CAS-9><CRISPR-based method><CRISPR-based technique><CRISPR-based technology><CRISPR-based tool><CRISPR/CAS approach><CRISPR/Cas method><CRISPR/Cas system><CRISPR/Cas technology><CRISPR/Cas9><CRISPR/Cas9 technology><Cas nuclease technology><Catabolism><Cell Body><Cells><Cellular Expansion><Cellular Growth><Cessation of life><Cholesterol><Clustered Regularly Interspaced Short Palindromic Repeats><Clustered Regularly Interspaced Short Palindromic Repeats approach><Clustered Regularly Interspaced Short Palindromic Repeats method><Clustered Regularly Interspaced Short Palindromic Repeats methodology><Clustered Regularly Interspaced Short Palindromic Repeats technique><Clustered Regularly Interspaced Short Palindromic Repeats technology><Codon><Codon Nucleotides><DNA><DNA Therapy><DNA cassette><Data><Death><Deoxyribonucleic Acid><Dialysis><Dialysis procedure><Disease><Disorder><Drugs><Ensure><Essential Genes><Essential Hypercholesterolemia><Failure><Familial Hypercholesterolemia><Family><Gene Targeting><Gene Transfer Clinical><Gene therapy trial><Genes><Genetic><Genetic Alteration><Genetic Change><Genetic Intervention><Genetic defect><Genome><Genomics><Goals><Grant><Hemapheresis><Hepatic Cells><Hepatic Disorder><Hepatic Parenchymal Cell><Hepatic Transplantation><Hepatocyte><Human><Hyperbetalipoproteinemia><Hypercholesteremia><Hyperlipemia><Hyperlipidemia><Hyperlipoproteinemia Type 2><Hyperlipoproteinemia Type II><Immune response><Immunological response><KO mice><Knock-out Mice><Knockout Mice><LDL><LDL Lipoproteins><LDL Receptors><LDLR gene><Length><Life><Lipoprotein LDL Receptors><Lipoproteins><Liver><Liver Cells><Liver Grafting><Liver Transplant><Liver diseases><Location><Low Density Lipoprotein Receptor><Low-Density Lipoproteins><Macaca><Macaque><Medication><Mendelian disease><Mendelian disorder><Mendelian genetic disorder><Mice><Mice Mammals><Modern Man><Morbidity><Morbidity - disease rate><Murine><Mus><Mutation><Null Mouse><Oncogenesis><Orphan Disease><Patients><Pharmaceutical Preparations><Pheresis><Physiology><Plasma><Plasma Serum><Procedures><Process><Programmed Cell Death><Protein Region><Proteins><QOL><Quality of life><Rare Diseases><Rare Disorder><Receptor Gene><Regenerative capacity><Reproducibility><Reticuloendothelial System, Serum, Plasma><Site><System><Technology><Testing><Therapeutic Gene Editing><Time><Transcription Regulation><Transcriptional Control><Transcriptional Regulation><Transgenes><Type 2 Hyperlipidemia><Type II Hyperlipidemia><Tyrosine><Viral><Work><Xanthomas><Zetia><adverse consequence><adverse outcome><atheromatosis><atherosclerotic disease><atherosclerotic vascular disease><autosome><beta-Lipoproteins><cell growth><curative intervention><curative therapeutic><curative therapy><curative treatments><dialysis therapy><dietary manipulation><drug/agent><enhancer cassette><expression cassette><ezetimib><ezetimibe><familial hyperbetalipoproteinemia><familial hypercholesteremia><familial hyperlipoproteinemia type 2><familial hyperlipoproteinemia type II><fatty liver disease><gene cassette><gene corrected><gene correction><gene editing method><gene editing methodology><gene editing strategy><gene editing techniques><gene repair therapy><gene therapy><gene transfer trial><gene-based therapy><gene-based treatment><gene-directed therapy><gene-editing approach><gene-editing therapy><gene-targeted therapy><gene-targeted treatment><genetic cassette><genetic therapy><genome editing><genome editing based therapy><genome editing therapy><genome editing treatment><genome editing-based therapeutics><genome mutation><genomic correction><genomic editing><genomic therapy><hepatic body system><hepatic disease><hepatic organ system><hepatopathy><high blood cholesterol><host response><human disease><hypercholesterolemia><immune system response><immunoresponse><in vivo><inhibitor><integration cassette><liver disorder><liver transplantation><loss of function mutation><mRNA Stability><monogenic disease><monogenic disorder><mortality><mouse model><murine model><native protein drug><new approaches><novel><novel approaches><novel strategies><novel strategy><orphan disorder><particle><pharmaceutical protein><pre-clinical><preclinical><preservation><promoter><promoter cassette><promotor><protein drug agent><protein-based drug><regeneration ability><regeneration capacity><repair><repaired><reporter cassette><resistance cassette><selectable cassette><selection cassette><single-gene disease><single-gene disorder><standard of care><stop cassette><therapeutic editing><therapeutic genome editing><therapeutic protein><therapeutic transgene><transcription cassette><transcriptional cassette><transgene><transgene cassette><tumorigenesis><uptake><western diet><western-style diet><western-type diet>