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Principal Investigator: Mats Ljungman
Organization: UNIVERSITY OF MICHIGAN AT ANN ARBOR
Fiscal Year: 2022
Award: $214,414
Funding agency: National Cancer Institute
ABSTRACT
Chromosome rearrangements are common in cancers and typically occur early in carcinogenesis.
They drive tumor development by altering the expression or function of oncogenes and tumor
suppressor genes by copy number alterations, formation of oncogenic fusions or by alterations of
DNA elements responsible for regulating cancer genes. Despite being a well-known feature of
cancer, discovered over hundred years ago, this common cancer hallmark has not yet been
therapeutically exploited. Now, for the first time, technological advancements in whole genome
sequencing and the development of CRISPR technologies for precision targeting makes it
possible to identify and target chromosome rearrangement junctions (CRJs) in tumor cells. In
addition, recent breakthroughs in in vivo delivery of CRIPR reagents involving nanoparticles will
open new avenues into clinical utilization. In this R21 application, we present a precision CRISPR
approach that aims at targeting cancer-specific CRJs as an untapped Achilles heel common to
all tumors. This CRISPR-based precision approach consists of pairs of CRJ-targeting guide RNAs
(gRNAs) that bring together two parts of a dCas9-conjugated endonuclease, Fok1, leading to its
activation and the induction of lethal DNA double strand breaks (DSBs) specifically in cancer cells.
Regardless of the tumor-driving mechanism, the presence of CRJs will be exploited to selectively
force these cancer cells to die. We have obtained proof-of-principle of the successful targeting of
CRJs with this approach in both colon and bladder cancer cell lines. We have assembled a multi-
disciplinary team with expertise in CRISPR technology and design, lipid nanoparticle delivery
systems and bladder cancer mouse models. With this team in place and with strong evidence of
efficient targeting of CRJs with Fok1-dCas9 in cancer cell lines, we are now primed to develop
and test our approach in pre-clinical cancer models using recently developed breakthrough lipid
nanoparticle delivery technologies. If successfully implemented, this paradigm-shifting precision-
targeting cancer therapeutic platform could be transformative in designing uniform treatments for
all cancer types irrespective of the biology of the tumor.
Terms: <Anti-Oncogenes><Antioncogenes><Automobile Driving><Benchmarking><Best Practice Analysis><Bioluminescence><Bladder Cancer><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><Cancer Genes><Cancer Induction><Cancer Model><Cancer Suppressor Genes><Cancer cell line><Cancer-Promoting Gene><CancerModel><Cancers><Cas nuclease technology><Cell Body><Cells><Chemistry><Chromosomal Synapsis><Chromosome Pairing><Chromosomes><Clinic><Clinical><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><Colon Cancer><Colon Carcinoma><Complex><DNA Alteration><DNA Double Strand Break><DNA Sequence Alteration><DNA mutation><Data Set><Dataset><Development><Drug Design><Drug Targeting><Drugs><Elements><Emerogenes><Encapsulated><Gene Copy Number><Gene Dosage><Generalized Growth><Generations><Genetic mutation><Growth><Guide RNA><Heterograft><Heterologous Transplantation><In Vitro><Malignant Bladder Neoplasm><Malignant Cell><Malignant Neoplasms><Malignant Tumor><Malignant Tumor of the Bladder><Malignant neoplasm of urinary bladder><Medication><Mice><Mice Mammals><Monitor><Murine><Mus><Non-Polyadenylated RNA><Onco-Suppressor Genes><Oncogenes><Oncogenes-Tumor Suppressors><Oncogenic><Pathway interactions><Personalized medical approach><Pharmaceutic Preparations><Pharmaceutical Preparations><Precision therapeutics><Proteins><RNA><RNA Gene Products><Reagent><Recessive Oncogenes><Ribonucleic Acid><Sequence Alteration><Site><Synapsis><System><Technology><Testing><Therapeutic><Time><Tissue Growth><Transforming Genes><Tumor Biology><Tumor Burden><Tumor Cell><Tumor Load><Tumor Suppressing Genes><Tumor Suppressor Genes><Urinary Bladder Cancer><Urinary Bladder Malignant Tumor><Xenograft><Xenograft procedure><Xenotransplantation><base><cancer cell><cancer in the colon><cancer type><carcinogenesis><design><designing><developmental><driving><drug/agent><endonuclease><entire genome><full genome><gRNA><genome sequencing><genomic alteration><in vivo><individualized approach><lipid based nanoparticle><lipid nanoparticle><malignancy><mouse model><multidisciplinary><murine model><nano particle><nano particle delivery><nano-sized particle><nanoparticle><nanoparticle delivered><nanoparticle delivery><nanosized particle><neoplasm/cancer><neoplastic cell><new approaches><novel><novel approaches><novel strategies><novel strategy><oncosuppressor gene><ontogeny><pathway><personalized approach><pre-clinical><precision approach><precision therapies><precision treatment><preclinical><protein complex><tailored approach><targeted cancer therapy><tumor><whole genome><xeno-transplant><xeno-transplantation>