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Principal Investigator: Hao Wu
Organization: BOSTON CHILDREN'S HOSPITAL
Fiscal Year: 2024
Award: $542,900
Funding agency: National Institute of Allergy and Infectious Diseases
Summary
A nanobody that recognizes immunoglobulin light chains, conjugated to a molecular entity that recognizes a
virus-infected or a cancerous cell, is an effective therapeutic: A single injection of fusion constructs comprising
an anti-kappa light chain nanobody (VHHkappa) and zanamivir, a small molecule that targets influenza
neuraminidase, protects mice from a lethal challenge with both A- and B-strains of influenza. In the model that
established protection by VHHkappa adducts against influenza, the underlying mechanism of action involves
antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC), but the
relative contribution of each is not known. We shall therefore use FcgR common g chain-deficient mice and C3-
deficient mice to assess the relative contributions of ADCC and CDC. The generation of Fc constructs of different
Ig isotypes and bearing FcR-engagement disabling mutations, similarly modified with zanamivir, will be used to
complement this analysis.
Having established proof-of-concept for influenza and optimized parameters for elimination of influenza virus-
infected cells, we will explore nanobodies that recognize other pathogens (Ebola virus, SARS-CoV-2, HIV) in
combination with VHHkappa in a series of collaborative experiments. The agents to be developed may inspire
novel immunomodulatory therapeutics, to be used as a stand-alone approach, or in combination with approved
drugs. The possibilities of post-exposure prophylaxis against viral infections (Ebola, SARS-CoV-2, HIV) in the
absence of pre-existing immunity, deserve particular emphasis. We shall further enhance the activity of the
proposed VHHkappa fusions through the generation of the corresponding drug adducts, using cytotoxic drugs such
as maytansinoids as compounds that have shown clinical utility. Enveloped viruses (e.g., HIV, SARS-CoV-2)
export viral proteins to the surface of the infected cell during budding. Infected cells can thus be distinguished
from uninfected cells based on the surface display of viral proteins.
We now extend these in vivo observations to fusions of VHHkappa with anti-checkpoint (PD-L1, CTLA-4)
nanobodies. We generated maytansinoid-modified VHHkappa fusions with the anti-PD-L1 and anti-CTLA-4 VHHs.
Our preliminary data show enhanced anti-tumor activity in the MC38 and B16.F10 mouse tumor models in
comparison with commonly used monoclonal antibodies. However, not all such fusions (examples: fusions of
VHHkappa with nanobodies that recognize Class II MHC or CD8) have shown the intended depletion efficacy in
vivo. This proposal seeks to establish the parameters -including biodistribution and surface expression levels of
the targeted molecules- that determine success or failure of VHHkappa fusions. The availability of VHHs that
recognize human kappa light chains suggest the possibility of clinical translation of this approach.
Terms: <2019 novel corona virus><2019 novel coronavirus><2019-nCoV><2019-nCoV S protein><2019-nCoV spike glycoprotein><2019-nCoV spike protein><4-guanidino-Neu5Ac2en><AIDS Virus><Ab-dependent cellular cytotoxicity><Acquired Immune Deficiency Syndrome Virus><Acquired Immunodeficiency Syndrome Virus><Acylneuraminyl hydrolase><Agonist><Allelism Test><Animals><Anti-Bacterial Agents><Anti-Infective Agents><Anti-Infective Drugs><Anti-Infectives><Anti-infective Preparation><Area><Assay><B7-H1><B7H1><Binding><Bioassay><Biodistribution><Biological Assay><Bone Marrow Purging><Boston><CD152><CD152 Antigen><CD152 Gene><CD274><CD8><CD8B><CD8B1><CD8B1 gene><COVID-19 S protein><COVID-19 spike><COVID-19 spike glycoprotein><COVID-19 spike protein><COVID-19 virus><COVID19 virus><CTLA 4><CTLA-4 Gene><CTLA4><CTLA4 gene><CTLA4-TM><Cancerous><Cell Body><Cells><Chimera Protein><Chimeric Proteins><Class II Genes><Clinical><Clinical Treatment Moab><CoV-2><CoV2><Colon Cancer><Colon Carcinoma><Complement-Dependent Cytotoxicity><Complementation Test><Cytotoxic T-Lymphocyte Protein 4><Cytotoxic T-Lymphocyte-Associated Antigen 4><Cytotoxic T-Lymphocyte-Associated Protein 4><Cytotoxic T-Lymphocyte-Associated Serine Esterase-4><Cytotoxic agent><Cytotoxic drug><Data><Disabling><Dose><Drugs><EBOV><Ebola><Ebola virus><Ebola-like Viruses><Failure><Fc Fragments><Fc Immunoglobulins><Fusion Protein><Generations><Genetic><Genetic Alteration><Genetic Change><Genetic Complementation Test><Genetic defect><Glycoproteins><Grippe><HIV><HIV Envelope Glycoprotein gp120><HIV Envelope Protein gp120><HIV env Protein gp120><HLA Class II Genes><HTLV-III gp120><Half-Life><Human><Human Immunodeficiency Viruses><IMiD><IgA><IgG1><IgG2><IgG3><IgG4><Immune Globulins><Immune modulatory therapeutic><Immunity><Immunoglobulin A><Immunoglobulins><Immunostimulatory drug><In Vitro><Influenza><Influenza Virus><Influenza Virus Infected Cells><Injections><Investigators><KO mice><Kappa Immunoglobulin Light Chain><Knock-out Mice><Knockout Mice><LAV-HTLV-III><LD-50><LD50><LYT3><Lead><Lethal Dose 50><Light><Light-Chain Immunoglobulins><Lymphadenopathy-Associated Virus><MC-38><MC38><MHC Class II><MHC Class II Genes><Malignant Melanoma><Mediating><Medication><Melanoma><Mice><Mice Mammals><Modeling><Modern Man><Molecular><Molecular Interaction><Monoclonal Antibodies><Murine><Mus><Mutation><N-Acylneuraminate Glycohydrolases><Neuraminidase><Neuraminidase inhibitor><Null Mouse><Oligosaccharide Sialidase><PD-L1><PDL-1><PDL1><Pb element><Phagocytosis><Pharmaceutical Preparations><Photoradiation><Programmed Cell Death 1 Ligand 1><Programmed Death Ligand 1><Prophylactic treatment><Prophylaxis><Publishing><Reaction><Relenza><Research Personnel><Researchers><SARS corona virus 2><SARS-CO-V2><SARS-COVID-2><SARS-CoV-2><SARS-CoV-2 S><SARS-CoV-2 S protein><SARS-CoV-2 spike><SARS-CoV-2 spike glycoprotein><SARS-CoV-2 spike protein><SARS-CoV2><SARS-associated corona virus 2><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-related corona virus 2><SARS-related coronavirus 2><SARSCoV2><Series><Severe Acute Respiratory Coronavirus 2><Severe Acute Respiratory Distress Syndrome CoV 2><Severe Acute Respiratory Distress Syndrome Corona Virus 2><Severe Acute Respiratory Distress Syndrome Coronavirus 2><Severe Acute Respiratory Syndrome CoV 2><Severe Acute Respiratory Syndrome-associated coronavirus 2><Severe Acute Respiratory Syndrome-related coronavirus 2><Severe acute respiratory syndrome associated corona virus 2><Severe acute respiratory syndrome coronavirus 2><Severe acute respiratory syndrome coronavirus 2 S protein><Severe acute respiratory syndrome coronavirus 2 spike glycoprotein><Severe acute respiratory syndrome coronavirus 2 spike protein><Severe acute respiratory syndrome related corona virus 2><Sialidase><Specificity><Surface><TLR7><TLR7 gene><Testing><Therapeutic Fungicides><Toll-Like Receptor 7><Trans Test><Transgenic Mice><Tumor Cell><Universities><Viral><Viral Diseases><Viral Gene Products><Viral Gene Proteins><Viral Proteins><Virus><Virus Diseases><Virus-HIV><Washington><Work><Wuhan coronavirus><aCTLA-4><aCTLA4><aPD-L1><aPDL1><adduct><anti programmed cell death ligand 1><anti programmed cell death protein ligand 1><anti-CTLA-4><anti-CTLA4><anti-PD-(L)1><anti-PD-L1><anti-PDL-1><anti-PDL1><anti-bacterial><anti-fungal><anti-fungal agents><anti-fungal drug><anti-tumor agent><antiPD-L1><antiPDL1><antibody dependent cell mediated cytotoxicity><antibody dependent cytotoxicity><antibody mediated cellular cytotoxicity><antibody-dependent cell cytotoxicity><antibody-dependent cellular cytotoxicity><antibody-mediated cytotoxicity><cancer in the colon><cancer microenvironment><clinical translation><clinically translatable><communicable disease control agent><complement-mediated cytotoxicity><complementation analysis><complementation approach><coronavirus disease 2019 S protein><coronavirus disease 2019 spike glycoprotein><coronavirus disease 2019 spike protein><coronavirus disease 2019 virus><coronavirus disease-19 virus><cytotoxic T-lymphocyte antigen 4><drug/agent><exo alpha sialidase><experiment><experimental research><experimental study><experiments><flu infection><flu serotype><flu strain><flu subtype><flu viral strain><flu virus infection><flu virus strain><genome mutation><gp120><gp120 ENV Glycoprotein><gp120(HIV)><hCoV19><heavy metal Pb><heavy metal lead><immune modulating agents><immune modulating drug><immune modulating therapeutics><immune modulatory agents><immune modulatory drugs><immune stimulatory agent><immune stimulatory drug><immune stimulatory therapeutic><immunomodulating agents><immunomodulating drugs><immunomodulator agent><immunomodulator drug><immunomodulator medication><immunomodulator prodrug><immunomodulator therapeutic><immunomodulatory agents><immunomodulatory drugs><immunomodulatory therapeutics><immunostimulatory agents><immunostimulatory biologics><improved><in vivo><in vivo Model><infected with flu><infected with flu virus><infected with influenza><infected with influenza virus><influenza infection><influenza serotype><influenza strain><influenza subtype><influenza viral strain><influenza virus infection><influenza virus strain><influenzavirus><kappa Immunoglobulins><kappa-Chain Immunoglobulins><mAbs><medical college><medical schools><monoclonal Abs><mutant><myeloablation><nCoV2><nanobodies><nanobody><neoplastic cell><novel><overexpress><overexpression><pathogen><pre-clinical study><preclinical study><programmed cell death ligand 1><programmed cell death protein ligand 1><protein death-ligand 1><receptor binding><receptor bound><recruit><response><school of medicine><sdAb><single domain antibodies><small molecule><sortase><spike proteins on SARS-CoV-2><srtA gene product><success><therapeutically effective><translation strategy><translational approach><translational strategy><tumor><tumor microenvironment><viral infection><virus infection><virus protein><virus-induced disease><zanamivir><α-CTLA-4><α-CTLA4><αCTLA-4><αCTLA4><αPD-L1><αPDL1>