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Principal Investigator: Arun Kannoth Nambiar
Organization: BOSTON UNIVERSITY (CHARLES RIVER CAMPUS)
Fiscal Year: 2024
Award: $40,671
Funding agency: National Cancer Institute
Project Summary Specific Aims
Solid tumors account for nearly 90% of adult cancers and are challenging to eliminate. Surgery and
adjuvant therapy (e.g., radiation, chemotherapy, and immunotherapy) are powerful methods for treating solid
tumors. Still, they are not universally effective or curative, and would benefit significantly from some means of
augmenting the tumor-killing response. Local delivery of cytotoxic, immunostimulatory, and gene therapy agents,
especially in combinations, has proven very effective in promoting solid tumor clearance. Given this potential, it
would be powerful to have a platform that sustainably and simultaneously delivers multiple immunostim-
ulatory and gene-therapy payloads directly at the tumor, thus promoting its clearance. While current
strategies, such as local drug delivery, lentivirus, and mRNA delivery methods, could generate such modifica-
tions, they suffer limitations for broader use such as permanent modifications and off-target effects (lentivirus),
transient gene expression (mRNA) and drug release, and limited payload. Self-amplifying RNA (saRNA) has
shown great promise for prolonged and non-integrative therapeutic gene expression and coupled with lipid na-
noparticles (LNPs) could serve as a viable method for local gene delivery and tumor modification. Until recently
the potent early immune response triggered upon entry of saRNA into the cell has severely constraining the
potency of saRNA, with standard strategies for lowering RNA immunogenicity such as incorporating the modified
nucleoside N1-methyl-pseudouridine (found in all mRNA COVID vaccine) yielding non-functional saRNA. How-
ever, we have recently discovered that complete substitution of saRNA with the 5 methylcytidine (5mC) drasti-
cally reduces immune recognition of saRNA while preserving saRNA replication and gene expression capabili-
ties, leading to elevated and prolonged transgene expression. This powerful discovery empowers the appli-
cation of saRNA for other therapeutic applications and provides an opportunity to optimize these thera-
pies for efficient, multiplex gene delivery. To achieve this, I propose studies for optimizing the saRNA platform
by two approaches. The first approach (aim 1) will involve screening both saRNA and LNP formulation to opti-
mize their gene delivery across a panel of tumor types, using our best-performing saRNA to express an innova-
tive gene drive cassette that would selectively enrich the expression of a suicide gene in the tumor and overcome
drug resistance. The second approach (Aim 2) will focus on maximizing transgene cargo in saRNA, screening
1) how many genes can be encoded in a single saRNA by implementing RNA sequence elements, and 2) the
maximum possible length of saRNA that can be successfully delivered. This work will establish new bench-
marks for RNA therapeutics, providing a platform for efficiently delivering the next generation of gene-
therapies for solid tumor treatment.
Terms: <2019-nCoV vaccine><21+ years old><Academia><Adjuvant Therapy><Adult><Adult Human><Benchmarking><Best Practice Analysis><Body Tissues><COVID-19 vaccine><Cancers><Cell Body><Cells><Chromosome Mapping><Communication><Coupled><Cytotoxic Chemotherapy><Cytotoxic Therapy><Cytotoxic agent><Cytotoxic drug><DNA Therapy><Data><Development><Drug Delivery><Drug Delivery Systems><Drug Precursors><Drug resistance><Drugs><EC 2.7.7.48><Elements><Enzyme Gene><Enzymes><Formulation><Gene Delivery><Gene Expression><Gene Localization><Gene Mapping><Gene Mapping Genetics><Gene Therapy Agent><Gene Transfer Clinical><Genes><Genetic Engineering><Genetic Engineering Biotechnology><Genetic Engineering Molecular Biology><Genetic Intervention><IRES><Immune><Immune mediated therapy><Immune response><Immunes><Immunological response><Immunologically Directed Therapy><Immunostimulatory drug><Immunotherapy><In Vitro><Industry><Internal Ribosome Entry Segment><Internal Ribosome Entry Site><Intratumoral heterogeneity><Investigators><Length><Lentivirinae><Lentivirus><Linkage Mapping><Lipids><Malignant Neoplasms><Malignant Tumor><Maps><Mediating><Medication><Mentors><Messenger RNA><Methods><Modification><Non-Polyadenylated RNA><Nucleosides><Operative Procedures><Operative Surgical Procedures><Pharmaceutical Preparations><Pro-Drugs><Prodrugs><Property><Proteins><Pseudouridine><R-Series Research Projects><R01 Mechanism><R01 Program><RNA><RNA Gene Products><RNA Replicase><RNA Sequences><RNA amplification><RNA based therapeutics><RNA based therapy><RNA replication><RNA therapy><RNA-Dependent RNA Polymerase><RNA-Directed RNA Polymerase><Radiation><Recombinant DNA Technology><Research><Research Grants><Research Personnel><Research Project Grants><Research Projects><Researchers><Ribonucleic Acid><Ribosome Entry Site><Ribosomes><SARS-CoV-2 vaccine><SARS-coronavirus-2 vaccine><Scientist><Severe Acute Respiratory Syndrome CoV 2 vaccine><Severe acute respiratory syndrome coronavirus 2 vaccine><Solid Neoplasm><Solid Tumor><Surgical><Surgical Interventions><Surgical Procedure><Technology><Testing><Therapeutic><Thinking><Tissues><Total Human and Non-Human Gene Mapping><Training><Transfection><Transgenes><Tumor Cell><Work><adjuvant treatment><adulthood><benchmark><cell type><chemotherapy><coronavirus disease 2019 vaccine><coronavirus disease-19 vaccine><cytokine><deliver mRNA><deliver messenger RNA><delivery system for mRNA><design><designing><developmental><drug resistant><drug/agent><empowerment><experience><experiment><experimental research><experimental study><experiments><gene repair therapy><gene therapeutics><gene therapy><gene-based therapeutic><gene-based therapeutics><gene-based therapy><genes therapeutic><genes therapeutics><genetic mapping><genetic payload><genetic therapy><genetically engineered><genomic therapy><heterogeneity in tumors><host response><immune stimulatory agent><immune stimulatory drug><immune stimulatory therapeutic><immune system response><immune therapeutic approach><immune therapeutic interventions><immune therapeutic regimens><immune therapeutic strategy><immune therapy><immune-based therapies><immune-based treatments><immuno therapy><immunogenic apoptosis><immunogenic cell death><immunogenicity><immunoresponse><immunostimulatory agents><immunostimulatory biologics><immunostimulatory therapy><immunostimulatory treatment><in vivo><innovate><innovation><innovative><interest><intra-tumoral heterogeneity><intratumor heterogeneity><lipid based nanoparticle><lipid nanoparticle><local drug delivery><mRNA><mRNA delivery><malignancy><materials science><messenger RNA delivery><multidisciplinary><nCoV vaccine><nCoV-19 vaccine><nCoV19 vaccine><nano particle><nano-sized particle><nanoparticle><nanosized particle><neoplasm/cancer><neoplastic cell><next generation><preservation><prevent relapse><promoter><promotor><relapse prevention><resistance to Drug><resistant to Drug><response><screening><screenings><skills><suicide gene><surgery><synthetic biology><therapeutic RNA><therapeutic gene><therapy optimization><thoughts><trafficking><transgene><transgene expression><treatment optimization><tumor><tumor heterogeneity><vaccine against 2019-nCov><vaccine against COVID-19><vaccine against SARS-CoV-2><vaccine against SARS-coronavirus-2><vaccine against Severe Acute Respiratory Syndrome CoV 2><vaccine against Severe acute respiratory syndrome coronavirus 2><vaccine candidates against SARS-CoV-2><vaccine for novel coronavirus><vaccines preventing COVID><vaccines to prevent COVID><vector>