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Principal Investigator: ELLIS L REINHERZ
Organization: DANA-FARBER CANCER INST
Fiscal Year: 2024
Award: $2,356,969
Funding agency: National Institute of Allergy and Infectious Diseases
SUMMARY – OVERALL
While there is an urgent need for vaccines eliciting broadly neutralizing antibodies (bnAbs) against the highly
mutable HIV-1 retrovirus to stem its global spread, this goal remains elusive. Antibodies produced against
trimeric gp160 sites of vulnerability during natural infection drive retroviral mutation further, diversifying quasi-
species in individuals. One apparent exception is the conserved membrane-proximal external region (MPER)
site, critical for hemifusion/fusion and which is stealth, largely immersed in lipid and only transiently revealed
during spontaneous ectodomain tilting. Liposome-arrayed MPER segments induce specific antibodies in mice
but without neutralizing activity, explained by the restricted antibody access to the native MPER residing in a
narrow “crawlspace” between the viral membrane below and the base of the gp120 and gp41 protomers of the
trimeric gp160 envelop. Biomaterial formulation of polymer “steric clouds” on liposomes or alternative origami-
based approaches on nanodiscs to limit access to the MPER, akin to that mandated by the trimer three-fold axis,
were technically cumbersome and confounded by the uncertainty of MPER topology. Our recent analysis of the
structure and dynamics of nanodisc-embedded trimeric HIV-1 spike protein provides compelling data for
alternative design. In addition, synthetic RNA lipid nanoparticle (LNP) encapsulation technology obviates vaccine
challenges with hydrophobic proteins such as the MPER and adjacent transmembrane segment (MPER-TM).
This P01 comprises two projects. Project 1 shall determine the immunogenicity of RNA vaccines encoding
MPER-TM trimers fused to structurally suitable self-proteins to focus antibody responses against the conjoint
membrane arrayed viral MPER-TM, with the self-proteins imposing steric restriction comparable to that exerted
by the gp160 ectodomain on the native MPER. Serological, single B cell, recombinant mAb methods,
neutralization assays and Ig bioinformatic analyses are applied. Automated computational searches of the pdb
informed by single-particle cryo-electron microscopy (cryo-EM) structures and using RNA technology reveal
antigenicity and immunogenicity of construct design for planned immunization studies in normal as well as knock-
in mice harboring human Ig D3-3 and JH6 gene segments for long CDRH3 loop generation. Project 2 will perform
cryo-EM and X-ray crystallographic studies to compare existing and additional HIV-1 patient-derived bnAbs
obtained from biobanked serial samples (Osier, IAVI) with those that are vaccine elicited, in the context of
nanodisc-arrayed MPER immunogens and native spike protein. We shall investigate Ig requirements of somatic
hypermutation (SHM), Fab molecular dynamics (MD) and co-evolution of antibody approach angles and affinities
of bnAbs of IgG1 and IgG3 isotypes. Projects 1 & 2 shall be performed by Kim/Weissman (DFCI/UPenn) and
Reinherz (DFCI), respectively, with four Technology Components: mRNA-LNP vaccine synthesis by Weissman
(U Penn); cryo-EM by Walz (Rockefeller), X-ray crystallography by Tan (APS) and Kwong (NIH); and MD by
Huang (Texas A&M). Administrative (Reinherz) and Antibody (Seaman, BIDMC) Cores serve all.
Terms: <7S Gamma Globulin><AIDS><AIDS Virus><Ab response><Acquired Immune Deficiency><Acquired Immune Deficiency Syndrome><Acquired Immune Deficiency Syndrome Virus><Acquired Immunodeficiency Syndrome><Acquired Immunodeficiency Syndrome Virus><Affinity><Antibodies><Antibody Formation><Antibody Production><Antibody Response><Antigen-Presenting Cells><Antigenic Determinants><Antigens><Assay><B blood cells><B cell><B cells><B-Cells><B-Lymphocytes><B-cell><Binding><Binding Determinants><Binding Sites><Bio-Informatics><Bioassay><Biocompatible Materials><Bioinformatics><Biological Assay><Biomaterials><Blood Plasma Cell><Bone Marrow><Bone Marrow Reticuloendothelial System><Charge><Chimera><Chimera organism><Clinical><Clinical Treatment Moab><Cloning><Combining Site><Complex><Cryo-electron Microscopy><Cryoelectron Microscopy><Data><Development><Electron Cryomicroscopy><Encapsulated><Envelope Protein><Epitopes><Face><Formulation><Fostering><Frequencies><Future><Generations><Genes><Genetic Alteration><Genetic Change><Genetic defect><Germ Lines><Glycans><Goals><Grain><HIV><HIV Envelope Glycoprotein gp120><HIV Envelope Protein gp120><HIV env Protein gp120><HIV-1><HIV-I><HIV1><HTLV-III gp120><Health><Human><Human Immunodeficiency Virus Type 1><Human Immunodeficiency Viruses><Human immunodeficiency virus 1><Hydrophobicity><Ig Genes><Ig Somatic Hypermutation><IgG><IgG1><IgG3><Immune system><Immunization><Immunoglobulin G><Immunoglobulin Genes><Immunoglobulin Somatic Hypermutation><In Vitro><Individual><Infection><KI mice><Knock-in Mouse><LAV-HTLV-III><Length><Licensing><Link><Lipid Binding><Lipids><Liposomal><Liposomes><Lymphadenopathy-Associated Virus><Mediating><Membrane><Messenger RNA><Methods><Mice><Mice Mammals><Modern Man><Modification><Molecular Dynamics Simulation><Molecular Interaction><Monoclonal Antibodies><Motion><Murine><Mus><Mutation><NIH><National Institutes of Health><Non-Polyadenylated RNA><Patients><Plasma Cells><Plasmacytes><Polymers><Polysaccharides><Preventative vaccine><Preventive vaccine><Principal Investigator><Prophylactic vaccine><Proteins><Proteome><Protomer><RNA><RNA Gene Products><RNA vaccine><RNA-based vaccine><Reactive Site><Recombinants><Retroviridae><Retroviruses><Ribonucleic Acid><Sampling><Serology><Single Crystal Diffraction><Site><Structure><Technology><Texas><Uncertainty><United States National Institutes of Health><Vaccines><Viral><Viral Diseases><Virus Diseases><Virus-HIV><Virus-Retrovirus><Writing><X Ray Crystallographies><X-Ray Crystallography><X-Ray Diffraction Crystallography><X-Ray/Neutron Crystallography><Xray Crystallography><accessory cell><antibody biosynthesis><arm><autoimmune reactivity><autoreactivity><base><bases><biobank><biological material><biorepository><chimeras><cryo-EM><cryoEM><cryogenic electron microscopy><design><design and construct><design and construction><designing><developmental><doubt><env Antigens><env Gene Products><env Polyproteins><env Protein><faces><facial><genome mutation><global health><gp120><gp120 ENV Glycoprotein><gp120(HIV)><gp160><immunization strategy><immunogen><immunogenicity><immunoglobulin biosynthesis><immunoreactivity><in vivo><knock-in animal><knockin animal><knockin mice><lipid based nanoparticle><lipid bound><lipid nanoparticle><mAbs><mRNA><mRNA vaccine><mRNA-based vaccine><membrane structure><molecular dynamics><monoclonal Abs><monoclonal antibody production><nanodisk><neutralizing antibody><particle><plasmocyte><polymer><polymeric><prevent><preventing><somatic hypermutation><stem><technology platform><technology system><tool><vaccination strategy><vaccination study><vaccination trial><vaccine formulation><vaccine study><vaccine trial><viral infection><virus infection><virus-induced disease>