2018 DARPA/EcoHealth 'DEFUSE' Proposal Documented All Three SARS-CoV-2 Spike Anomalies Before the Pandemic
Furin cleavage site, human-optimized RBD, and the two-proline spike mutation all appear—together—in EcoHealth’s pre-pandemic DEFUSE proposal to DARPA.
Summary:
A 2018 EcoHealth Alliance DARPA proposal, “DEFUSE,” outlined plans to engineer and aerosolize chimeric SARS-related coronaviruses and self-spreading vaccines using large-area spray systems and drones.
A USRTK FOIA release produced 1,417 pages of early DEFUSE drafts and internal emails from U.S. Geological Survey records.
Those documents show that all three major structural anomalies of the SARS-CoV-2 spike—furin cleavage site insertion, human-optimized receptor-binding domain mutations, and the two-proline (2P) spike-stabilizing substitution—were explicitly described in DEFUSE before the pandemic.
While each of these features has been treated as a separate unexplained phenomenon, DEFUSE is the only known pre-pandemic proposal documenting all three in one place.
The DEFUSE plans match the precise spike architecture later found in SARS-CoV-2 and the stabilized spike used in mRNA vaccines.
DEFUSE is EcoHealth Alliance’s 2018 Defense Advanced Research Projects Agency (DARPA) proposal for a joint U.S.–China program to engineer and aerosolize chimeric SARS-related coronaviruses, immune modulators, and self-spreading vaccines—even delivered via large-area spray systems and drones for “inoculation of animals/humans.”
A Freedom of Information Act (FOIA) request by the investigative nonprofit public health research group ‘U.S. Right to Know’ (USRTK) led to the release of a large batch of documents related to DEFUSE.
USRTK’s FOIA release included a 1,417-page file of documents obtained from the U.S. Geological Survey that contained early drafts of the DEFUSE proposal submitted to DARPA and related emails from individuals on the project.
Significantly, that file reveals that all three of the greatest mysteries surrounding the COVID-19 spike protein—furin cleavage site (FCS) insertion, receptor-binding domain (RBD) mutations, and two-proline (2P) substitutions—were explicitly characterized in DEFUSE.
While all three of these spike-centric mysteries have individually been the subject of many reports at various levels of analysis, to our knowledge, no single report has made it clear that all three of them are simultaneously documented in the DEFUSE proposal.
Among more than 100 catalogued natural sarbecoviruses, not one contains even a single one of these anomalies—yet EcoHealth’s DEFUSE proposal lists all three together in 2018.
Each anomaly is independently unprecedented in nature, and DEFUSE remains the only pre-pandemic document in existence to describe all three. The coincidence is statistically and historically extraordinary.
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Furin Cleavage Site in DEFUSE
The furin cleavage site mystery is that SARS-CoV-2, the COVID-19 pandemic pathogen, is the only known sarbecovirus in nature to possess a four-amino acid insertion “PRRA” at the S1/S2 junction that creates a perfect furin-cleavable motif.
The PRRA insertion creates a furin cleavage site that enables the SARS-CoV-2 spike protein to be efficiently cut and activated by the human enzyme furin, dramatically increasing viral entry into cells, tissue tropism, and transmissibility compared to any previously known natural coronavirus.
This insertion, never previously observed in any of its hundreds of known relatives, is explicitly proposed for addition into coronavirus spike proteins in the DEFUSE project.
Pages 524–525 of USRTK’s DEFUSE document outline plans to “introduce a furin cleavage site” into pre-pandemic coronavirus chimeras around amino acid residues 667–792.
The unique SARS-CoV-2 pandemic spike FCS amino acid motif PRRA begins at residue 681, fitting perfectly within reach of the 667–797 residues designated in DEFUSE in pre-pandemic strains.
A February 2022 Frontiers in Virology study noted that the 12-nt PRRA insertion sits within a 19-nucleotide sequence that is a 100% match to the reverse complement of a codon-optimized mRNA construct patented by Moderna (US 9,587,003), a pattern the authors describe as “highly unusual” and absent from all other mammalian or viral genomes.
The optimized SARS-CoV-2 furin cleavage site appears in DEFUSE.
Receptor-Binding Domain in DEFUSE
The receptor-binding domain mystery is that SARS-CoV-2’s RBD is uniquely optimized for human ACE-2 binding through a precise set of 14 contact residues confirmed by Ralph Baric of the University of North Carolina at Chapel Hill.
These residues include the most interaction-dense ACE-2 interaction hotspot Q498 (encoded by the rare ‘CAA’ glutamine codon), which anchors key ACE-2 sites D38 and K353 while stabilizing the RBD internally.
The closest known purportedly natural relative, BANAL-52, matches SARS-CoV-2 at 13 of these 14 residues, differing only at position 498 (histidine instead of glutamine), the exact site pre-pandemic gain-of-function research by Baric mutated to glutamine, and which DEFUSE explicitly proposed targeting for human-optimization mutations.
DEFUSE page 524 outlines plans to mutate coronavirus spike proteins at residues 442, 479, 487, and 491 with deletions at 432–437 and 485–472.
DEFUSE document page 522 outlines plans for “human-optimized RBD residues” at residue 442, 472, 479, 480, and 487.
Ralph Baric published a chart in a March 2020 Journal of Virology paper confirming all of these pre-pandemic residues, and their SARS-CoV-2 equivalents, are critical for spike protein binding to human ACE-2 receptors.
A pandemic-era BioEssays analysis noted that SARS-CoV-2 possesses both a furin cleavage site “missing in other CoVs of the same group” and an RBD “optimized to bind to human cells,” and concluded that the simultaneous acquisition of these features is “less likely to be natural” and “might be the result of lab manipula-tion techniques such as site-directed mutagenesis.”
The humanized SARS-CoV-2 receptor-binding domain appears in DEFUSE.
‘2P’ (Two-Proline) in DEFUSE
The two-proline (2P) mystery is that the exact double-proline substitution (V1060P + L1061P in SARS-CoV-1 numbering) explicitly proposed in the 2018 DEFUSE proposal as a deliberate engineering step to stabilize novel bat coronavirus spikes in prefusion conformation—an exact modification Baric’s team had already tested in pre-pandemic strains—was later incorporated into the stabilized spike proteins used in the Pfizer and Moderna mRNA vaccines (K986P + V987P), even though the naturally circulating SARS-CoV-2 virus retains the ancestral KV motif found in every other known sarbecovirus.
In other words: COVID-19 mRNA vaccines contain the exact 2P engineering step that DEFUSE openly planned to insert into coronavirus spikes, while the naturally circulating SARS-CoV-2 virus carries the precise KV amino acid residue sequence that DEFUSE had already targeted for that proline replacement.
DEFUSE document page 399 outlines plans to add “two proline residues at positions V1060P and L1061P [to] stabilize the prefusion state of the trimer, including key neutralizing epitopes in the receptor binding domain.”
Astonishingly, relative target locations to V1060 and L1061 in the spike protein encoded by Pfizer Inc. and Moderna Inc. mRNA vaccines (K986 and V987) are replaced with the same two proline amino acids in order to “stabilize the resulting spike protein in the prefusion state and contribute to vaccine efficiency,” according to a January 2021 Viruses publication.
Baric collaborators performed gain-of-function experiments to specifically mutate V1060 and L1061 in MERS-CoV, thereby introducing two prolines, as reported in an August 2017 PNAS publication.
The spike-stabilizing two-proline substitution appears in DEFUSE.
Bottom Line
The DEFUSE documents show that every major structural anomaly that distinguishes SARS-CoV-2 from all known natural sarbecoviruses—its furin cleavage site, its human-optimized receptor-binding domain, and its Pfizer/Moderna-linked two-proline (2P) spike-stabilizing mutations—was pre-specified in a 2018 U.S.–China engineering proposal submitted by EcoHealth Alliance, Ralph Baric, and the Wuhan Institute of Virology.
Each of these three features has been treated for years as an independent “mystery” requiring elaborate natural-origin explanations (praeter Austria’s Segreto et Canada’s Deigin).
Yet DEFUSE reveals that all three were simultaneously planned, described, and targeted before the pandemic by the very actors later positioned at the center of the global outbreak response.
This alignment between DEFUSE’s engineering plans and SARS-CoV-2’s defining spike features represents the strongest circumstantial evidence to date that the COVID-19 pandemic virus’s spike architecture matches the blueprint of a specific, documented, pre-pandemic laboratory program.
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Exceptional investigative work pulling these USRTK documents together. The statistical convergence you document is what makes this so compeling: each individual anomaly has been debated endlessly, but the fact that DEFUSE explicitly describes all three modifications in a single proposal before they appeared in SARS-CoV-2 transforms this from circumstantial evidence into something much harder to dismiss. What's particularly striking is how the 2P modification ended up in the mRNA vaccines but not the circulating virus, which basically means vaccine manufacturers independently arrived at the exact same stabilzation strategy DEFUSE had already outlined for gain of function work.
It was all there, and people like you and Alina Chan and Matt Ridley and Kevin McKernan have helped get it out--but you'll still never find it reported in MSM! WE HAVE A LONG WAY TO GO!