4/1/2020

PROTOTYPE APPROACH TO PANDEMIC PREPAREDNESS

HIV→PNEUMOVIRUS→PARAMYXOVIRUS→CORONAVIRUS

ADVAC Alumni 2 April 2020

Barney S. Graham, MD, PhD Deputy Director , NIAID, NIH 1

Recent Zoonotic and Vector-borne Viral Threats

• Hanta • Nipah/Hendra • West Nile virus • SARS • • • • MERS • Zika • EV‐D68 • SARS‐CoV‐2

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NIAID Vaccine Research Center

Basic Research Process Development Nucleic acid

Vectors

VLPs

• AIDS/HIV • Influenza Proteins and cGMP Manufacturing nanoparticles • Ebola/Marburg • RSV • Malaria Monoclonal antibodies • Tuberculosis • EID GLP Analysis • West Nile virus, Zika • Chikungunya • W/E/V equine encephalitis • MERS‐CoV, SARS, and other CoV • Nipah and other paramyxoviruses • EV‐D68 and other picornaviruses Clinical Trials • Smallpox

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Public health burden of re-emerging & emerging viruses

Traditional Approaches

• Licensed vaccines/antibiotics Vaccine • Passive surveillance Challenges • Contact tracing • Quarantine • Vaccines for unmet needs • Emerging viruses • Improving licensed vaccines

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New Technologies are Transforming Vaccinology

• Structure-based vaccine design Structural analysis of antigenic sites on viral • Single-cell sorting, sequencing, and bioinformatics surface glycoproteins – Rapid isolation of human mAbs Isolation of human monoclonal – Definition of antibody lineages antibodies from single B cells – Analysis of immune responses • Protein engineering of self-assembling nanoparticles • Rapid DNA synthesis

• Recombinant DNA and genetic engineering technology Epitope-specific phenotyping – Rapid cell line development Sequencing for viral diversity and escape mutations – Animal model development • Nucleic acid and vector-based delivery of vaccine

Sequencing B cells to define clonal lineages; TCR & BCR-specific transcriptome 5 5

New Technologies Facilitate an Engineering Approach

Vaccine New Challenges Technologies

• Vaccines for • Structural biology unmet needs • Protein engineering • Emerging viruses • Single cell sorting and analysis • Improving • High throughput sequencing licensed vaccines • Rapid isolation of human mAbs • Antibody lineage analysis • Rapid diagnostic tools • Systems biology

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Gene Synthesis and Platform Technologies

Historical knowledge about virus Isolation of virus and extraction …ATGCGACT… biology and structure. and sequencing of genome Understanding antigenic targets Digital transfer of and basis for immunity information

Surveillance and Discovery Design and Synthesis

Virus‐infected person from Synthetic Vaccinology Synthesis of key genes encoding new outbreak vaccine and other reagents

Graham & Sullivan. Immunology 2018 7 7

Platform Technologies Shorten Manufacturing Timelines

100 days → ~50 days

Graham, Mascola, Fauci. Novel Vaccine Technologies: Essential Components of an Adequate Response to Emerging Viral Diseases JAMA2018 8 8

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Structure-guided Stabilization of HKU1 CoV Spike

V1060P L1061P

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2-P Mutation Stabilizes MERS and SARS CoV S Improved expression, prefusion structure, immunogenicity

50‐fold greater expression than WT

MERS S‐2P ‐ V1060P & L1061P

MERS S‐2P stabilizes prefusion trimers by EM

Prefusion S Postfusion S

Pallesen, J., Wang, N., Corbett, K., … Graham BS, Ward A, McLellan et. al. PNAS 2017 14

2P mutations effectively stabilize multiple CoV prefusion S

Pallesen, J.*, Wang, N.*, Corbett, K.*, et. al. PNAS. 2017. 15

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2P mutations effectively stabilize multiple CoV prefusion S

Potentially Endemic Human CoVs Porcine CoV Emerging CoV

EM Reconstruction by Robert Kirchdoerfer (Ward lab | Scripps) 16

Immunogenicity of MERS S-2P in Mice

Pallesen, J., Wang, N., Corbett, K., et. al. PNAS 2017 | EM by Yaroslav Tsybovsky 17 17

Stabilized MERS Spike-2P Protects hDPP4 Knock-in Mice from Lethal Challenge

Adam Cockrell, Ralph Baric, 18 18

6 4/1/2020

MERS S-2P mRNA elicits better neutralizing antibodies than MERS S-WT mRNA

Dose Titration 0.016 g Immunogen Dose MERS S-2P vs. MERS S WT mRNA **** 106 105

105 4 **** 10 Titer Titer 50 4 50 10 103 103 S WT 102 102

S-2P Reciprocal IC Reciprocal IC 1 1 10 10 S WT S-2P 0.01 0.1 1 Immunogen Dose (µg) Immunonogen Group

Dr. Kizzmekia Corbett in collaboration with Therapeutics

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Summary

• S from multiple CoV strains can be stabilized in the prefusion conformation using homologous 2P mutations

• Stabilized prefusion S trimers are more immunogenic and protective than WT trimers or monomeric subunits

• May be a general solution for beta‐CoV vaccine antigen design

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CORONAVIRUS BIOLOGY AND NOMENCLATURE

corona = crown or circle of light

Spike Protein

Wrapp D, Wang N, Corbett KS, Goldsmith JA, Hsieh CL, Abiona O, Graham BS, McLellan JS. Cryo‐EM structure of the 2019‐nCoV spike in the prefusion conformation. Science. 2020 Feb 19:eabb2507. doi: 10.1126/science.abb2507.

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Vaccine Target: Coronavirus Spike Protein • Coronavirus spike protein is on the viral surface and mediates attachment to cells to start the infection process • Ideal vaccines target coronavirus spikes in order to block viral infection

Spike Protein Human ACE2 Receptor

Coronavirus CellsCells inin HumanHuman BodyBody

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Vaccine Target: Coronavirus Spike Protein • Coronavirus spike protein attaches to ACE2 protein to start infection

Coronavirus CellsCells inin HumanHuman BodyBody

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Vaccine Target: Coronavirus Spike Protein • Vaccine‐induced antibodies will block the interaction and function of CoV spike protein

Antibodies

Coronavirus CellsCells inin HumanHuman BodyBody

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COVID-19 VACCINE DEVELOPMENT

2013-2019 2020 2021 SARS-CoV-2 Outbreak

2013-2019 Jan 6, 2020 Jan 11, 2020 Jan 14, 2020 Jan 31, 2020 Feb 4, 2020 Feb 10, 2020 Feb 19, 2020 Feb 21, 2020 March 16, 2020

Spike Extensive work Wuhan 2019-nCoV Moderna VRC makes VRC UT-Austin Moderna Phase 1 Clinical structure on MERS and outbreak may constructs receives nCoV spike vaccinates solves spike ships vaccine trial Starts published in other CoV be CoV ordered sequence for protein mice structure to DMID Science GMP production

Dec 31, 2019 Jan 10, 2020 Jan 13, 2020 Jan 23, 2020 Feb 1, 2020 Feb 7, 2020 Feb 18, 2020 Feb 20, 2020 Mar 2, 2020

1st report of 2019-nCoV VRC decides DMID First nCoV Moderna Immunogenicity DMID FDA safe-to- respiratory virus sequences on vaccine initiates spike ELISA vials vaccine confirmed in submits IND proceed outbreak in published sequence clinical and for cross- mice Wuhan, China with Moderna regulatory reactivity process

3 days 38 days 24 days

Previous fastest timeline 90 days 100 days ZIKV DNA vaccine

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COVID-19 Vaccine Consortium

NIAID OD: NIAID VRC USG organization Vaccine design Vanderbilt Communications Assay development Live virus PRNT Diplomatic Relations Preclinical evaluation Human immunology UW IPD Pathogenesis NIAID DMID: Nanoparticle display IND VTEU clinical trial sites Advanced development UT Austin Spike structure NIAID DIR RML animal NHP model

UNC: Industry Molecular clone CDC Non‐profit and mouse model COVID‐19 Virus isolation Vaccine Diagnostics Government Sample sharing Public health organizations Development WHO Academia Multiple Advisory and WG committees and US Dept of State coordination NIAID office of Global Affairs Negotiations with WHO and China

CEPI DOD WRAIR: Funding Prior MERS study and collaborators

Abcellera FDA CBER: mAb discovery Regulatory oversight and BARDA/ASPR: advice Moderna Development support, USG GMP mRNA organizational leadership

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Structure of Prefusion RSV F Glycoprotein

Prefusion Postfusion

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Generalized Application to Other Fusion Proteins

Graham, Gilman, McLellan, Annual Review of Medicine 2019

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Summary

• New technologies are transforming vaccinology providing solutions for long-standing problems and emerging viral diseases • Combining atomic level antigen design with computationally designed nanoparticles and platform manufacturing approaches provides a modular, engineering approach to achieve generalizable solutions for vaccine antigen design • The advent of precision vaccinology with rapid platform manufacturing makes a prototype pathogen approach for pandemic preparedness feasible

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NIAID Vaccine Research Center

Viral Pathogenesis Laboratory VPL Tracy Ruckwardt Masaru Kanekiyo Kaitlyn Morabito Michelle Crank Kizzmekia Corbett Seyhan Boyoglu‐Barnum Lauren Chang Rebecca Gillespie Emily Phung Adrian Creanga Man Chen Syed Moin Deepika Nair Julia Lederhofer Olubukola Abiona Geoffrey Hutchinson Azad Kumar Brian Fisher Alex Derrien‐Coleman Cynthia Ziwawo Anthony Dipiazza Osnat Rosen Rebecca Loomis Karin Bok Erez Bar Haim Saavan Chintalacheruvu Monique Young LaTanya Chapman

VRC NIAID Julie Ledgerwood & Clinical Trial Program John Mascola Mario Roederer Anthony Fauci Adrian McDermott & Vaccine Immunology Program Richard Koup Daniel Douek Hilary Marston Diane Scorpio & Animal Care Program Sarah Andrews Robert Seder Cristina Cassetti Jason Gall & Vaccine Production Program Peter Kwong Nancy Sullivan Audray Harris David Lindsay & Vaccine Clinical Material Program Jeffrey Boyington Judy Stein/Will Adams Theodore Pierson Kevin Carlton & Project Management Abe Mittelman Marybeth Daucher

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Acknowledgements

VRC Immunology Cores Kathy Foulds Amy Noe Dillon Flebbe Nadesh Nji Shing-Fen Kao Valerie Ficca Madhu Prabhakaran UT Austin Scripps Neil King Jason McLellan Andrew Ward Joshua Brand David Baker David Ambrozak Morgan Gilman Jesper Pallesen Daniel Ellis Funding:DB, NPK Brooke Fiala Nianshuang Wang Robert Kirchdoerfer Strategic Planning & Development George Ueda Mike Battles Christopher Cottrell Jason Gall Jorge Fallas Daniel Wrapp Hannah Turner Judith Stein Rashmi Ravichandran Will Adams Lauren Carter Vanderbilt UNC Kevin Carlton Mythreyi Shastri Mark Denison Ralph Baric Jim Chappel Adam Cockrell

Yaroslav Tsybovsky Tyler Stephens

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NIAID VRC Clinical Trials Program

Ingelise Sarah Martin Grace Chen Gordon Plummer Gaudinski Deputy Chief Clinical Chief, Clinical Medical Operations Development Director Manager Unit Charla Andrews Aba Eshun Lam Le Stephanie Taylor Preeti Apte Carmencita Graves Floreliz Mendoza Cora Trelles Cartagena Julie Ledgerwood Alison Beck Mercy Guech Laura Novik Olga Trofymenko Chief, Clinical Trials Program Eugeania Burch Cynthia Starr Hendel Mark O’Callahan Olga Vasilenko Maria Burgos Florez Somia Hickman Abidemi Ola Xiaolin Wang Nina Berkowitz Cristina Carter Renunda Hicks Iris Pittman Wil Whalen Team Lead, Emily Coates LaSonji Holman Ro Rothwell Alicia Widge Protocol Operations Pam Costner Kate Houser Jamie Saunders Pernell Williams Josephine Cox Rebecca Lampley Ellie Seo Galina Yamshchikov Jennifer Cunningham Brenda Larkin Sandra Sitar

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