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Principal Investigator: Venigalla B. Rao
Organization: CATHOLIC UNIVERSITY OF AMERICA
Fiscal Year: 2024
Award: $964,530
Funding agency: National Institute on Drug Abuse
This proposal aims to establish a new category of safe and affordable in vivo HIV cure “drugs” that would
transform the health of substance users who are disproportionately affected by HIV-1 (HIV) disease. Substance
users suffer from high rates of HIV acquisition and transmission, poor adherence to antiretrovirals and drug
resistance, and high prevalence of neurodegenerative and mental health problems. It will also be broadly
transformative to more than 40 million people across the globe who are living with HIV today.
Using a unique bacteriophage (phage) T4 artificial viral vector (T4-AVV) technology we have recently
developed, we will engineer the 120 x 86 nm T4 capsid nanoparticle to target patient's hematopoietic stem cells
(HSCs) in vivo and deliver a payload of genome modification molecules. These will introduce a delta-32 deletion
mutation into the CCR5 HIV co-receptor gene, which will make the HSCs and the cells derived from it, HIV-
resistant. This is the only known HIV cure option today, as evident from at least five independent HIV cure cases
worldwide, all remarkably involving transplanted HSCs containing the naturally present delta-32 mutation.
Our experimental path involves three milestones. First, we will identify targeting ligands that have high affinity
and binding specificity to HSCs from human umbilical cord blood (UCB). These will be selected from libraries of
nanobodies, darpins, and ScFvs displayed on phage T4 capsid through fusion at the tip of the 180Å-long,
flexible, outer fibers known as Hoc. Next, HSC-targeting, lipid-coated, T4-AVVs will be assembled in the test
tube by incorporating all the HIV cure molecules into one nanoparticle by sequential assembly and DNA
packaging. These include combinations of genome editing enzymes, gRNAs, and donor DNAs. The T4-AVVs
that exhibit maximum delta-32 genome conversion will be optimized using an ex vivo UCB-HSC model. Finally,
the HSC-targeted T4-AVVs will be tested in vivo in a humanized DRAGA mouse model reconstituted with the
same UCB-HSCs, using a protocol that mimics HIV cure in humans. Various parameters will be optimized such
that the T4-AVVs will enter HSCs in minutes, deliver payload in hours, and modify genome in 1-2 days.
The T4-AVV in vivo HIV-cure would be a safe, affordable, outpatient protocol that can be easily administered
to substance users. Since it is targeted, a small dose of T4-AVVs (~10E12 particles) would be sufficient to
capture the HSCs mobilized into patient's blood, while minimizing host immune responses and off-target events.
The delta-32 converted HSCs will then repopulate the body with HIV-resistant blood cells including CD4+ T cells,
which would replace the HIV reservoir and result in functional HIV cure. A single dose of T4-AVVs might be
sufficient, however a second dose may be needed to maximize HSC delta-32 conversion for permanent HIV
cure. Currently, T4-AVV is the only technology that has the capacity and engineering capability to create such
HIV cure delivery vehicles. Furthermore, this plug-and-play drug model can be adapted to many other blood cell
diseases and cancer, potentially transforming future gene therapies and precision medicine.
Terms: <AIDS Virus><Acquired Immune Deficiency Syndrome Virus><Acquired Immunodeficiency Syndrome Virus><Adherence><Affect><Affinity><Anti-retroviral drug resistance><Bacteriophage T4><Bacteriophages><Binding><Blood><Blood Cells><Blood Precursor Cell><Blood Reticuloendothelial System><C-C CKR-5><C-C CKR-5 Gene><C-C Chemokine Receptor Type 5><C-C Chemokine Receptor Type 5 Gene><CC Chemokine Receptor 5><CC-CKR-5><CC-CKR-5 Gene><CC-CKR5><CCCKR5><CCCKR5 Gene><CCR-5><CCR-5 Gene><CCR5><CCR5 Protein><CCR5 Receptors><CCR5 gene><CD195 Antigen><CD195 Antigen Gene><CD4 Cells><CD4 Positive T Lymphocytes><CD4 T cells><CD4 helper T cell><CD4 lymphocyte><CD4+ T-Lymphocyte><CD4-Positive Lymphocytes><CHEMR13><CHEMR13 Gene><CKR-5><CKR-5 Gene><CKR5><CKR5 Gene><CKR5 Receptors><CMKBR5><CMKBR5 Gene><Cancers><Capsid><Categories><Cell Body><Cells><Chemokine (C-C Motif) Receptor 5><Chemokine (C-C) Receptor 5><Chemokine (C-C) Receptor 5 Gene><Coliphage T4><Cord Blood><Cord Blood Hematopoietic progenitor><Cord Blood Hematopoietic stem cells><DNA><DNA Packaging><DNA Therapy><Deletion Mutation><Deoxyribonucleic Acid><Disease><Disorder><Dose><Drug Modelings><Drugs><Engineering><Enterobacteria phage T4><Enzyme Gene><Enzymes><Event><Exhibits><Fiber><Future><Gene Transfer Clinical><Genetic><Genetic Alteration><Genetic Change><Genetic Intervention><Genetic defect><Genome><Guide RNA><HIV><HIV Infections><HIV resistance><HIV resistant><HIV-1><HIV-1 Fusion Co-Receptor><HIV-1 Fusion Co-Receptor Gene><HIV-I><HIV1><HSC transplantation><HTLV-III Infections><HTLV-III-LAV Infections><Health><Hematopoietic Progenitor Cells><Hematopoietic Stem Cell Mobilization><Hematopoietic Stem Cell Transplant><Hematopoietic Stem Cell Transplantation><Hematopoietic progenitor Mobilization><Hematopoietic progenitor cell Mobilization><Hematopoietic stem cells><High Prevalence><Hour><Human><Human Immunodeficiency Virus Type 1><Human Immunodeficiency Viruses><Human T-Lymphotropic Virus Type III Infections><Human immunodeficiency virus 1><Immune response><Immunological response><In vivo analysis><Infusion><Infusion procedures><LAV-HTLV-III><Libraries><Ligands><Lipids><Lymphadenopathy-Associated Virus><Malignant Neoplasms><Malignant Tumor><Medication><Mental Health><Mental Hygiene><Modern Man><Modification><Molecular Interaction><Mutation><Nerve Degeneration><Neuron Degeneration><Out-patients><Outpatients><Patients><Peripheral Blood Cell><Persons><Phages><Pharmaceutical Preparations><Play><Population><Progenitor Cells><Protocol><Protocols documentation><Psychological Health><Receptor Gene><Resistance><Specificity><T-Cells><T-Lymphocyte><T4 Cells><T4 Lymphocytes><T4 Phage><Technology><Testing><Transmission><Tube><Umbilical Cord Blood><Vaccines><Viral Vector><Virus><Virus-HIV><anti-retroviral drug resistant><bacterial virus><blood cell progenitor><blood progenitor><blood stem cell><blood-forming stem cell><delivery vector><delivery vehicle><drug/agent><fetal cord blood><flexibility><flexible><gRNA><gene repair therapy><gene therapy><gene-based therapy><gene-based treatment><gene-directed therapy><gene-targeted therapy><gene-targeted treatment><genetic therapy><genome editing><genome mutation><genomic editing><genomic therapy><hematopoietic cell transplantation><hematopoietic cellular transplantation><hematopoietic progenitor><hematopoietic progenitor cell transplantation><hematopoietic stem and progenitor cell mobilization><hematopoietic stem progenitor cell><hemopoietic progenitor><hemopoietic stem cell><host response><human progenitor><human stem cells><immune system response><immunoresponse><in vivo><in vivo evaluation><in vivo testing><infusions><malignancy><mouse model><murine model><nano particle><nano-sized particle><nanobodies><nanobody><nanoparticle><nanosized particle><neoplasm/cancer><neural degeneration><neurodegeneration><neurodegenerative><neurological degeneration><neuronal degeneration><novel><particle><precision medicine><precision-based medicine><progenitor cell model><progenitor model><reconstitute><reconstitution><resistance to anti-retroviral drug><resistant><resistant to anti-retroviral drug><sdAb><single domain antibodies><stem and progenitor cell model><stem cell based model><stem cell derived model><stem cell model><stem cells><substance user><thymus derived lymphocyte><transmission process><vector>