13484SUBS
THE OVERALL GOAL OF THE OREGON HEALTH & SCIENCE UNIVERSITY (OHSU) PROGRAM IS TO INFORM THE RATIONAL DESIGN OF AN EFFECTIVE TB VACCINE BY IDENTIFICATION OF IMMUNE RESPONSES THAT ARE CAPABLE OF CONTROLLING OR ERADICATING MTB. THE HUMAN IMMUNOLOGIC STUDIES (LEAD BY DR. LEWINSOHN AND HIS TEAM) WILL FOCUS ON THE FOLLOWING ACTIVITIES IN THIS PERIOD: HOUSEHOLD CONTACTS STUDY Â ¢ SUB-ACTIVITY 2 (AIM 1): PERFORM SCRNASEQ ON NON-ADHERENT LYMPHOCYTES TO DEFINE THE PHENOTYPIC AND FUNCTIONAL PROFILE OF NON-MYELOID CELLS IN THE LUNGS AND PERIPHERAL CIRCULATION. SUB-ACTIVITY 5 (AIM 4): PREPARE A MINIMUM OF 100 TCR CLONES OF EACH STUDY PARTICIPANT OF ALL COHORTS AND EVALUATE FOR HLA-RESTRICTION, FUNCTIONAL PHENOTYPE, TCR USAGE, AND ANTIGENIC SPECIFICITY
Oregon Health & Science University
Definitive Contract 75N93019C00070
$410.1k 1/31/25 13488SUBS
THE DU PLESSIS/WALZL LABORATORY AT STELLENBOSCH UNIVERSITY (SU) WILL BE RESPONSIBLE FOR THE CLINICAL AND IMMUNOLOGICAL WORK (INCLUDING ON-SITE DATA MANAGEMENT) OF THE PROJECT. SU WILL COORDINATE THE STUDY DESIGN, PARTICIPANT SCREENING AND ENROLLMENT FOR THE CASCADE HOUSEHOLD CONTACT STUDY, PERFORM BRONCHOSCOPIES AND MANAGE SAMPLE SHARING TO PARTICIPATING SITES. SU WILL PROVIDE INPUT IN THE DESIGN, EXECUTION AND DATA ANALYSIS OF IMMUNOLOGICAL STUDIES AND PARTICIPATE IN THE LEADERSHIP TEAM OF Â CASCADE IMPAC-TBÂ CENTER. SU WILL INTERACT AND COORDINATE ALL PROJECT ACTIVITIES WITH THE PI OF THE PROJECT. DR. DU PLESSIS AND DR. WALZL, THE SU CO-PIÂ S WILL REVIEW ALL RAW DATA GENERATED BY THE SU TEAM, AND SUPERVISE THE LABORATORY WORK, AND THE RESEARCH TECHNICIANS. SPECIFIC DELIVERABLES FOR YEAR 6 OF THE CONTRACT AT SU INCLUDE THE FOLLOWING: A. RECRUITMENT: THE ANTICIPATED SCREENING AND RECRUITMENT GOALS HAVE BEEN ACHIEVED AND EXCEEDED. WHILE THE INITIAL RECRUITMENT TARGET WAS SET AT 20 PARTICIPANTS FOR EACH OF THE 8 CLINICAL GROUPS (N=160 TOTAL), WE EXCEEDED THIS TO ACHIEVE AND ENROLL A TOTAL NUMBER OF 185 STUDY PARTICIPANTS ACROSS ALL GROUPS FOR THE MAIN STUDY. THE COVID-19 PANDEMIC HAS RESULTED IN A REQUEST FOR A SOMEWHAT ALTERED CLASSIFICATION AND ENROLLMENT TARGET OF CLINICAL GROUPS, WHICH WAS APPROVED BY THE NIH IN 2021. THE SU HHC STUDY NOW CONSISTS OF GROUPS STRATIFIED NOT ONLY BY IGRA RESULTS OR PET/CT READOUT, BUT ALSO BY SARS-COV-2 SEROLOGY STATUS. THE BUDGET FOR PERIOD 6 WILL INCLUDE LIMITED PARTICIPANT RECRUITMENT AND BRONCHOSCOPIES, AS ORIGINALLY PROPOSED, WHICH WILL INCLUDE ONLY STUDY PARTICIPANTS FROM SELECTED Â BIOLOGICALLY INTERESTINGÂ CLINICAL PHENOTYPES FOR FURTHER TARGETED FUNCTIONAL ANALYSES THROUGH GRANULOMA MODEL MANIPULATION. SU HAS BUDGETED FOR N=12 BRONCHOSCOPIES. B. END-ASSAYS: IMMUNOPHENOTYPING EXPERIMENTS Â ONE OF THE DELIVERABLES AT SU IS TO CONDUCT DEEP PHENOTYPING OF PAIRED BRONCHOALVEOLAR LAVAGE (BAL) AND PBMC FROM THE VARIOUS CASCADE HOUSEHOLD CONTACT STUDY CLINICAL GROUPS. SU HAS EVALUATED THE ASSAY PRIORITY LIST, AS WELL AS THE BAL AND PBMC SAMPLES COLLECTED FROM PARTICIPANTS ENROLLED IN THE CASCADE HOUSEHOLD CONTACT STUDY, TO ENUMERATE THE NUMBER OF STORED SAMPLES AVAILABLE FOR THE OBJECTIVES RELATED TO THE PROPOSED PLANS FOR THIS END-ASSAY. OF ALL MAIN STUDY PARTICIPANTS FROM WHICH PAIRED BAL AND PBMC HAVE BEEN COLLECTED (N=190), SUFFICIENT CELL NUMBERS WERE OBTAINED FROM 88% OF PARTICIPANTS, WHICH INCLUDES MAKING USE OF RESIDUALLY STORED BAL CELLS. THIS ALSO INCLUDES REPLACEMENT OR REPEATING OF BRONCHOSCOPIES FOR N=5 OF THE PARTICIPANTS WITH LOW CELL YIELDS. WHILE THE BAL CELL AUTOFLUORESCENCE IS LIKELY TO RESULT IN A SMALLER NUMBER OF AVAILABLE FLUORESCENT CHANNELS FOR OUR BAL MYELOID FLOW CYTOMETRY IMMUNE PHENOTYPING PANEL, THE OPTIMIZATION OF THE 24 MARKER MYELOID PANEL FOR PBMC HAS BEEN COMPLETED WHILE OPTIMIZATION OF THE 27 MARKER LYMPHOID PANEL FOR PBMC WILL BE FINALIZED IN OCTOBER 2024. SU EXPECTS TO COMPLETE IMMUNE PHENOTYPING OF ALL SU HHC PBMC SAMPLES BY JUNE 2025. IMMUNE PHENOTYPING OF ALL SU HHC BAL SAMPLES USING THE LYMPHOID FLOW CYTOMETRY PANEL WILL BE COMPLETED BY DECEMBER 2025. GRANULOMA MODEL/KILLING ASSAYS Â SU WILL MAKE USE OF THE IN VITRO GRANULOMA MODEL ALONG WITH PARALLEL ASSESSMENTS IN MONOLAYER CULTURE AND UNFRACTIONATED PBMC AND BAL, TO ASSESS MTB KILLING AMONGST THE VARIOUS CLINICAL GROUPS. INTERESTING FINDINGS RELATED TO MTB CONTROL WILL BE TESTED IN FUTURE IN GRANULOMA MODEL EXPERIMENTS. BY USING CLINICAL SAMPLES FROM PARTICIPANTS NOT ELIGIBLE ACCORDING TO THE CASCADE HHC STUDY CRITERIA, SU HAS OPTIMIZED THE MODEL AND ALSO IMPLEMENTED CHANGES TO STREAMLINE BACTERIAL LOAD ASSESSMENT THROUGH MGIT CULTURE. SU EXPECTS TO COMPLETE THE KILLING ASSAY EXPERIMENTS FOR AT LEAST A QUARTER OF ALL SU HHC SAMPLES BY THE END OF PERIOD 6. THE RESULTS FROM VARIOUS CASCADE OMICS READOUTS (HUMAN AND LAB ANIMAL RESULTS) WILL INFORM FUTURE GRANULOMA MODEL MANIPULATIONS TO TEST THE FUNC
Stellenbosch University
Definitive Contract 75N93019C00070
$613.1k 1/31/25 13492SUBS
STATEMENT OF WORK: DEFINING THE ROLE OF CD8 T CELLS INDUCED BY A NOVEL LIPOSOMAL NANOPARTICLE MRNA VACCINE IN PREVENTING MYCOBACTERIUM TUBERCULOSIS INFECTION. THE GOAL OF IMPAC-TB IS TO IDENTIFY MECHANISMS OF PROTECTIVE IMMUNITY AGAINST TB, AND OUR OVERALL STRATEGY FOR THE CASCADE IMPAC-TB ANIMAL PROGRAM IS TO START WITH GOLD-STANDARDS OF VACCINE-MEDIATED PROTECTION (I.E, CMV/TB AND IV BCG IN NHP; BCG IN MICE) AND THEN DISSECT HOW PROTECTION IS MEDIATED. THE AKAGERA MRNA VACCINE REPRESENTS A NEW GOLD STANDARD FOR PREVENTION OF INFECTION Â THE BEST VACCINE-MEDIATED PROTECTION EVER OBSERVED IN MICE. TO ACHIEVE OUR GOAL OF IDENTIFYING MECHANISMS OF TB IMMUNITY, CASCADE IMPAC-TB WOULD LIKE TO IDENTIFY THE ANTIGENS AND T CELL SUBSETS THAT MEDIATE THE PROTECTION PROVIDED BY THE NEW AKAGERA MRNA VACCINE, WHICH SHOWS THE BEST VACCINE-MEDIATED PROTECTION EVER OBSERVED IN MICE. THE SPECIFIC WORK TO BE PERFORMED IS DETAILED BELOW. UNDER ALL THREE AIMS, AKAGERA MEDICINES WILL BE RESPONSIBLE FOR CREATING THE MRNA VACCINE CONSTRUCTS REQUIRED FOR EACH EXPERIMENT. IN ADDITION, SCIENTISTS AT AKAGERA MEDICINES WILL ALSO MEET REGULARLY WITH SCIENTISTS AT SCRI TO EVALUATE PROGRESS, DESIGN NEW CONSTRUCTS, AND TROUBLE-SHOOT, AS NECESSARY. AIM 1: IDENTIFY PROTECTIVE ANTIGENS IN THE AKAGERA MRNA VACCINE. AKAGERA MEDICINES WILL PREPARE SEPARATE MRNA VACCINE CONSTRUCTS FOR 4 ORFS (TB10.4, MTB39, AND ESAT-6 WHICH ARE THE MOST IMMUNOGENIC ANTIGENS IN THE MRNA VACCINE, AND MTB32, WHICH ALONG WITH MTB39 IS PART OF THE M72 VACCINE). THESE WILL BE SENT TO SCRI FOR TESTING IN MICE, TO COMPARE THE PROTECTIVE EFFICACY OF THE PARENTAL MRNA VACCINE, EACH OF THESE 4 CONSTRUCTS INDIVIDUALLY, POOLED ADMINISTRATION OF TB10.4, MTB39, ESAT-6 MRNA CONSTRUCTS, AND THAT SAME POOL WITH THE ADDITION OF MTB32. SCRI WILL MEASURE THE THREE METRICS OF IMMUNITY THAT WERE PREVIOUSLY DEFINED IN THE ULD MODEL: REDUCTION OF LUNG BACTERIAL BURDENS, PREVENTION OF DISSEMINATION TO THE CONTRALATERAL LUNG, AND PREVENTION OF DETECTABLE INFECTION. COLONIES WILL BE COLLECTED, DNA ISOLATED, AND BAR-CODES AMPLIFIED AND SEQUENCED TO DETERMINE THE NUMBER OF FOUNDING MTB STRAINS IN EACH LUNG. BECAUSE THE GROUP SIZES WILL HAVE TO BE REDUCED TO 15 MICE (RATHER THAN OUR MORE STANDARD 20) TO ACCOMMODATE THE NUMBER OF VACCINES TESTED SIMULTANEOUSLY, THE EXPERIMENT WILL BE REPEATED FOUR TIMES TO ASSURE REPRODUCIBILITY AND STATISTICAL RIGOR. IF PROTECTION CONFERRED BY THE THREE OR FOUR POOLED ANTIGENS IS NOT SIMILAR TO THAT CONFERRED BY THE PARENT VACCINE, AKAGERA MEDICINES WILL SYNTHESIZE ADDITIONAL ANTIGENS UNTIL A COCKTAIL OF ANTIGENS THAT CONFERS SIMILAR PROTECTION IS FOUND. AIM 2: IDENTIFY AND ELIMINATE CD8 EPITOPES FROM MRNA VACCINE CONSTRUCTS. THIS AIM WILL FOCUS ON EACH ANTIGEN THAT CONFERRED SOME DEGREE OF PROTECTION AGAINST ESTABLISHING SUSTAINED INFECTION IN AIM 1. ANTIGENS THAT DID NOT PREVENT DETECTABLE INFECTION WILL NOT BE EXAMINED FURTHER IN AIM 2. IN PRIOR CD8 EPITOPE MAPPING STUDIES, TB10.4 IS KNOWN TO HAVE TWO EPITOPES, TB10.4 ON THE H-2B BACKGROUND (C57BL/6) AND TB10.4 ON THE H-2D BACKGROUND (BALB/C), AND MTB32A IS KNOWN TO HAVE ONE EPITOPE ON THE H-2B BACKGROUND (C57BL/6) (MTB32). THUS, IF THESE ANTIGENS ARE FOUND TO CONTRIBUTE TO PREVENTION OF INFECTION IN AIM 1, AKAGERA MEDICINES WILL MAKE A TRUNCATED TB10.4 OR MTB32A CONSTRUCT THAT LACKS THE RESPECTIVE CD8 EPITOPES, BUT WHICH WOULD STILL CONTAIN KNOWN CD4 EPITOPES. FOR MTB39 AND ESAT6, THE CD8 EPITOPES HAVE NOT BEEN MAPPED, BUT IF EITHER MTB39 OR ESAT-6 WERE SHOWN IN AIM 1 TO CONTRIBUTE TO PREVENTING INFECTION, AKAGERA WOULD GENERATE OVERLAPPING PEPTIDES FOR EACH ANTIGEN AND MAP THE EPITOPE(S) RESPONSIBLE USING STIMULATIONS ASSAYS WITH SPLENOCYTES FROM IMMUNIZED B6XBALB/C F1 MICE. AFTER IDENTIFYING THE EPITOPES, AKAGERA MEDICINES WOULD GENERATE NEW CONSTRUCTS THAT SPECIFICALLY LACKED THE CD8 EPITOPES, EITHER VIA TRUNCATION OR POINT MUTATION OF AMINO ACID RESIDUES PREDICTED TO BE CRITICAL FOR THE RESPONSE. MICE WOULD BE I
Akagera Medicines Inc.
Definitive Contract 75N93019C00070
$107.3k 1/31/25 13490SUBS
THE PURPOSE OF THIS SUBAWARD IS TO STUDY HOW THE BASELINE STATE AND FUNCTION OF ALVEOLAR MACROPHAGES CAN BE ALTERED BY VACCINATION AND OTHER HOST FACTORS, IN THE CONTEXT OF MYCOBACTERIUM TUBERCULOSIS INFECTION. THE PROPOSED STUDIES WILL INVESTIGATE THE MECHANISMS THAT DRIVE INNATE CELL REPROGRAMMING BY VACCINATION AND ITS IMPACT ON THE HOST IMMUNE RESPONSE AND BACTERIAL CONTROL. THE SUBAWARD WILL ALSO PERFORM A CROSS-SPECIES ANALYSIS OF ALVEOLAR MACROPHAGES BY ANALYZING EPIGENETIC STATES ACROSS MOUSE AND HUMAN SAMPLES AND LINKING MACROPHAGE EPIGENETIC STATES TO TRANSCRIPTIONAL RESPONSES. 1) ANALYZE EPIGENETIC PROFILES GENERATED BY ATAC-SEQ AND TRANSCRIPTIONAL PROFILES GENERATED BY RNA-SEQ FOR UNTREATED VERSUS INTERFERON-TRAINED MURINE ALVEOLAR MACROPHAGES. 2) PERFORM ATAC-SEQ EPIGENETIC PROFILING ON FROZEN HUMAN ADHERENT BAL SAMPLES FROM ACTIVE TB, RESISTER, HEALTHY CONTROLS, AND SUBCLINICAL TB INDIVIDUALS. PROFILING INCLUDES: CELL PREPARATION, TRANSPOSITION REACTION AND PURIFICATION, PCR AMPLIFICATION, LIBRARY PURIFICATION, AND LIBRARY QC. 3) COMPARE AND CONTRAST MOUSE AND HUMAN ALVEOLAR MACROPHAGE TRANSCRIPTIONAL AND EPIGENETIC PROFILES. 4) GENERATE BULK RNA-SEQUENCING SAMPLES FOR MTB-INFECTED B6 AND C3H ALVEOLAR MACROPHAGES FOLLOWING SA161 HIGH DOSE AEROSOL INFECTION. 5) PERFORM IN VITRO MTB INFECTIONS AND IN VIVO AEROSOL INFECTIONS AFTER AM TRANSFER USING WT, SP140-/-, AND SP140-/-IFNAR-/- MURINE EX VIVO ALVEOLAR MACROPHAGE (MEXAM) PRIMARY CELL LINES TO STUDY ALVEOLAR MACROPHAGE RESPONSE TO VACCINATION AND INTERFERON EXPOSURES.
University Of Massachusetts
Definitive Contract 75N93019C00070
$108.7k 1/31/25 13485SUBS
UNDER THIS OPTION WE WILL CONDUCT STUDIES IN RM TO VALIDATE THE EFFICACY OF THE RHCMV/TB VECTOR ORTHOLOG OF THE HUMAN CMV VACCINE CANDIDATE (VIR-1778) CURRENTLY SCHEDULED FOR TESTING IN A PHASE 1 CLINICAL TRIAL. WE WILL ALSO DEVELOP AND TEST RHCMV VECTORS THAT ARE MODIFIED TO REDUCE OR ELIMINATE THE DELETERIOUS IMMUNOREGULATORY EFFECTS OF THE CONTROL RHCMV VECTORS THAT LED TO ACCELERATED TB DISEASE, IN PARTICULAR VECTOR BACKBONE MODIFICATIONS THAT WILL KEEP THE RHCMV/TB VECTOR OUT OF THE LUNG, AS WELL AS OBVIATE ITS ABILITY TO INFECT MONOCYTE/MACROPHAGES, ENDOTHELIAL AND EPITHELIAL CELLS SAND TO PRODUCE IMMUNOREGULATORY VIRAL (V) IL-10 (YEARS 6-7). SUB-ACTIVITY 1: GENERATE A RHCMV/TB (AND RHCMV/CONTROL) VECTOR BASED ON THE VIR-1778 HCMV/TB VECTOR (FL-RHCMV + Â RH157.5-Â RH157.4 + RH107 R MTB-7AG) AND 4 DIFFERENT MODIFIED VERSIONS OF THIS ORTHOLOG VECTOR DESIGNED TO OBVIATE THE DELETERIOUS IMMUNOREGULATION MENTIONED ABOVE. ALL VECTORS WILL BE MADE AS BOTH TB-7AG INSERT EXPRESSION VECTORS (USING THE SAME TB-7AG CASSETTE BEING USED IN VIR 1778), AND AS A CONTROL VECTOR EXPRESSING AN INSERT OF ROUGHLY SIMILAR SIZE EXPRESSING UNRELATED SIV AGS. SUB-ACTIVITY 2: SELECT 2 OF THE 4 BACKBONE DESIGNS FROM SUB-ACTIVITY 1 AND THE VIR-1778 ORTHOLOG VECTOR SET FOR IMMUNOGENICITY AND EFFICACY TESTING IN MONKEYS, COMPARING THE BACKBONES WITH MTB VS. CONTROL AG INSERTS VS. UNVACCINATED CONTROLS. THE FOLLOWING GROUPS WILL BE TESTED USING UP TO A TOTAL OF 56 MONKEYS: Â ¢ VECTOR A (VIR-1778 ORTHOLOG) AND MATCHING CONTROL AG INSERT VECTOR; 8 MONKEYS EACH Â ¢ VECTOR B* (MODIFIED VIR-1388 ORTHOLOG VECTOR #1) AND MATCHING CONTROL AG INSERT VECTOR; 7-8 MONKEYS EACH Â ¢ VECTOR C* (MODIFIED VIR-1388 ORTHOLOG VECTOR #2) AND MATCHING CONTROL AG INSERT; 7-8 MONKEYS EACH, AND Â ¢ 8 UNVACCINATED CONTROLS SUB-ACTIVITY 3: PERFORM TIMED NECROPSIES AT WEEK 4 OR 5 POST-CHALLENGE AND PERFORM MICROBIOLOGIC AND HISTOPATHOLOGIC/IN SITU ANALYSES TO MEASURE EXTENT OF MTB INFECTION. SUB-ACTIVITY 4: CONDUCT LONGITUDINAL AND END-POINT (NECROPSY) ASSAYS TO MONITOR TRANSCRIPTOMIC PREDICTORS OF VACCINE EFFICACY AND THE DEVELOPMENT AND RESPONSE TO MTB CHALLENGE OF MTB-SPECIFIC CD4+ AND CD8+ T CELL RESPONSES AND INNATE Â TRAINEDÂ IMMUNITY. VECTOR B AND C TO BE SELECTED FROM VECTORS WITH THE FOLLOWING ADDITIONAL MODIFICATIONS (THAT IS ADDED TO TO THE VIR-1378 ORTHOLOG VECTOR BACKBONE) Â ¢ SINGLE CYCLE VERSION: + RH108-FKBP Â ¢ VIL-10 DELETED: + Â RH143 Â ¢ MYELOID, ENDOTHELIAL AND EPITHELIAL TROPISM-RESTRICTED AND VIL-10 DELETED: + Â RH143 + MIR126/142/205 (RH108/RH156) Â ¢ DOUBLE-DELETED AND MYELOID, ENDOTHELIAL AND EPITHELIAL TROPISM-RESTRICTED AND VIL-10 DELETED: + Â RH143 + MIR126/142/205 (RH108/RH156) + Â RH158-Â RH161
Oregon Health & Science University
Definitive Contract 75N93019C00070
$8.0m 1/31/25