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Principal Investigator: SANJAY RAM
Organization: UNIV OF MASSACHUSETTS MED SCH WORCESTER
Fiscal Year: 2024
Award: $208,392
Funding agency: National Institute of Allergy and Infectious Diseases
PROJECT SUMMARY/ABSTRACT
About 87 million new cases of gonorrhea occur worldwide annually. In 2020, 677,769 cases were reported to the CDC, a
111% increase in annual incidence since the historic low in 2009. Neisseria gonorrhoeae (Ng) has become resistant to
almost every antibiotic in clinical use. Reports of resistance to ceftriaxone – the only currently recommended first-line of
treatment – from almost every continent portends an era of untreatable gonorrhea. Development of novel treatments
and preventives against Ng is a global public health priority.
Our group, in collaboration with Evaxion Biotech, has identified two putative cell-division proteins (NGO0265 and
NGO1549 (FtsN)) as lead vaccine candidates. NGO0265 and FtsN are expressed by all Ng isolates and FtsN is essential for
bacterial viability. Immunization of mice with NGO0265 and FtsN elicits bactericidal antibodies (Abs) and significantly
reduces the duration and burden of gonococcal colonization of mouse vaginas. Ng deploy a unique immune evasion
strategy wherein it caps its lipooligosaccharide (LOS) with sialic acid using its surface LOS sialyltransferase (Lst) and host-
derived CMP-sialic acid. LOS sialylation enables Ng to evade complement, cationic antimicrobial peptides (CAMPs) and
down-regulate the inflammatory response by engaging host Siglec receptors. Gonococcal lst deletion mutants are
attenuated in mice. Supporting its key role in pathogenesis, all gonococcal disease isolates sequenced thus far possess a
functional Lst gene.
Camelid single-domain antibodies called VHHs or nanobodies, have been found to possess unique properties that offer
enormous versatility to facilitate the rapid development of simple and economical antibody-based therapeutics. VHH-
based products can exploit the small size and high stability of VHH components, their propensity to bind conformation-
dependent neutralizing epitopes, and their tractability for functional expression as heteromultimers. These characteristics
make possible commercially favorable antibody therapeutic agents having ultrahigh target affinities, broad natural variant
specificities and the ability to bind multiple different targets.
Here, we aim to develop nanobody-based therapeutics against NGO0265, FtsN and Lst for use as an adjunctive treatment
and as a preventive against multidrug-resistant gonorrhea. In Aim 1, we will immunize alpacas with purified recombinant
chimeric NGO0265-NGO1549 and Lst, create a nanobody-display phage library from immune B cells and identify
nanobodies that recognize non-overlapping epitopes on their targets with high-affinity. The ability of the nanobodies to
block enzyme function or to activate complement and kill Ng when linked to IgG Fc will be evaluated in Aim 2. In Aim 3,
we will test the ability of the lead anti-Ng nanobodies against each of the targets to clear gonococcal colonization in a
well-established mouse vaginal colonization model.
Successful completion of the work will identify and characterize nanobodies with activity against multidrug-resistant Ng,
which can then be adapted to a variety of formats and drug delivery platforms.
Terms: <7S Gamma Globulin><AIDS Virus><Acquired Immune Deficiency Syndrome Virus><Acquired Immunodeficiency Syndrome Virus><Affect><Affinity><Alpaca><Anti-microbial Cationic Peptides><Antibiotic Agents><Antibiotic Drugs><Antibiotic Resistance><Antibiotics><Antibodies><Antibody Therapy><Antigenic Determinants><Antigens><Antimicrobial Cationic Peptides><Attenuated><B blood cells><B cell><B cells><B-Cells><B-Lymphocytes><B-cell><Bacteria><Binding><Binding Determinants><Binding Proteins><Biotech><Biotechnology><C 5b-9><C5b-9><CMP Acetylneuraminic Acid><CMP-NANA><CMP-Sialic Acid><Cannot achieve a pregnancy><Cefatriaxone><Ceftriaxone><Cell Body><Cell division><Cells><Characteristics><Chlamydia><Clinical><Collaborations><Complement><Complement Complex C5b-9><Complement Factor H><Complement Membrane Attack Complex><Complement Proteins><Cytidine Monophosphate N-Acetylneuraminic Acid><Cytolytic Terminal Complement Complex><Development><Difficulty conceiving><Disease><Disorder><Drug Delivery><Drug Delivery Systems><Drug Resistance in N. Gonorrhoeae><Drug resistance in Neisseria Gonorrhoeae><Drug resistant N. Gonorrhoeae><Drug-resistant N. Gonorrhoeae><Drug-resistant Neisseria Gonorrhoeae><Ectopic Pregnancy><Enzyme Gene><Enzymes><Epitopes><Factor H><Flow Cytofluorometries><Flow Cytofluorometry><Flow Cytometry><Flow Microfluorimetry><Flow Microfluorometry><Genes><Genetic Alteration><Genetic Change><Genetic defect><Goals><Gonococcal Infection><Gonococcus><Gonorrhea><HIV><Human><Human Immunodeficiency Viruses><IgG><IgG1><Immune><Immune Evasion><Immunes><Immunization><Immunize><Immunoglobulin G><Immunotherapeutic agent><In Vitro><Incidence><Individual><Infection><Infertility><Inflammatory Response><LAV-HTLV-III><Lead><Libraries><Ligand Binding Protein><Ligand Binding Protein Gene><Link><Lymphadenopathy-Associated Virus><Mediating><Membrane Attack Complex><Messenger RNA><Mice><Mice Mammals><Microbe><Microbicidal Cationic Proteins><Miscellaneous Antibiotic><Miyagawanella><Modeling><Modern Man><Molecular Configuration><Molecular Conformation><Molecular Interaction><Molecular Stereochemistry><Multi-Drug Resistance><Multidrug Resistance><Multiple Drug Resistance><Multiple Drug Resistant><Murine><Mus><Mutation><N gonorrhea><N gonorrhoeae><N-Acetylneuraminic Acids><N. gonorrhea><N. gonorrhoeae><Neisseria gonorrhea><Neisseria gonorrhoeae><Neisseria gonorrhoeae antibiotic resistance><Organism><Pathogenesis><Pathway interactions><Pb element><Persons><Phage Display><Predisposition><Prevention><Preventive><Property><Protein Binding><Proteins><Reaction><Receptor Protein><Recombinants><Recommendation><Reporting><Resistance><Resistance to Multi-drug><Resistance to Multidrug><Resistance to Multiple Drug><Resistance to antibiotics><Resistant to Multiple Drug><Resistant to antibiotics><Resistant to multi-drug><Resistant to multidrug><Sexually Transmitted Diseases><Sexually Transmitted Disorder><Sexually Transmitted Infection><Sialic Acids><Sialyltransferases><Specificity><Surface><Susceptibility><Technology><Terminal Complement Complex><Testing><Therapeutic><Therapeutic Agents><Therapeutic antibodies><Topical Drug Administration><Topical application><Transmission><United States><Vagina><Vaginal Ring><Variant><Variation><Venereal Diseases><Venereal Disorders><Venereal Infections><Virus-HIV><Woman><Work><administer topically><anti-toxin><antibiotic drug resistance><antibiotic resistant><antibiotic resistant N. gonorrhoeae><antibiotic resistant Neisseria gonorrhoeae><antibiotic resistant gonorrhea><antibiotic-resistant N. gonorrhoeae><antibody based therapies><antibody treatment><antibody-based therapeutics><antibody-based treatment><apply topically><attenuate><attenuates><bactericidal><bactericide><bedsonia><bound protein><burden of infection><chronic pelvic pain><chronic pelvic pain syndrome><complementation><conformation><conformational><conformational state><conformationally><conformations><deliver topically><developmental><drug-resistant gonorrhea><efficacy testing><extrauterine pregnancy><fertility cessation><fertility loss><flow cytophotometry><genome mutation><gonococcal antibiotic resistance><heavy metal Pb><heavy metal lead><hyperimmunization><immune clearance><immune drugs><immune elimination><immune evasive><immune-based therapeutics><immunogen><immunologic therapeutics><immunotherapeutics><immunotherapy agent><improved><in silico><in vivo><infection burden><infertile><inhibitor><intravaginal ring><lipid-linked oligosaccharides><lipooligosaccharide><living system><mRNA><mucosal site><multi-drug resistant><multidrug resistant><mutant><nanobodies><nanobody><novel><pathway><prevent><preventing><public health priorities><receptor><resistant><screening><screenings><sdAb><sexually acquired infection><sialic acid binding Ig-like lectin><sialylation><siglec><single domain antibodies><synergism><technology platform><technology system><topical administration><topical delivery><topical drug application><topical treatment><topically administered><topically applied><topically delivered><topically treated><transmission process><treat topically><trend><vaccine candidate>