Records Show EcoHealth/Wuhan Research to Create Coronavirus ‘Mutants’


 

All reports submitted by EcoHealth Alliance to NIH or its sub-agencies related to NIH Grant No. 1R01A|110964 titled “Understanding the Risk of Bat Coronavirus Emergence” during the term of the grant.

To understand the risk of zoonotic CoV [coronavirus] emergence, we propose to examine 1) the transmission dynamics of bat-CoVs across the human-wildlife interface; and 2) how this process is affected by CoV evolutionary potential, and how it might force CoV evolution. We will assess the nature and frequency of contact among animals and people in two critical human-animal interfaces: live animal markets in China and people who are highly exposed to bats in rural China.

We will test our models of host range (i.e. emergence potential) experimentally using reverse genetics, pseudovirus and receptor binding assays, and virus infection experiments in cell culture and humanized mice. With bat-CoVs that we’ve isolated or sequenced, and using live virus or pseudovirus infection in cells of different origin or expressing different receptor molecules, we will assess potential for each isolated virus and those with receptor binding site sequence to spill over. We will do this by sequencing the spike (or other receptor binding/fusion) protein genes from all our bat-CoVs, creating mutants to identify how significantly each would need to evolve to use ACE2, CD26/DPP4 (MERS-CoV receptor) or other potential CoV receptors.

In vitro [outside the body] cell lines & Humanized mouse model: We have developed primary cell lines and transformed cell lines from 9 bat species using kidney, spleen, heart, brain and intestine. We have used these for virus isolation, infection assays and receptor molecule gene cloning. We also have a large number of cell lines from humans and animals that we will use for virus infectivity assays. We have obtained a letter of support from Dr Ralph Baric, who is keen to collaborate with us initially to infect his humanized mouse model with our bat SL-CoV [SARS-Like Coronavirus] that uses ACE2, and subsequently to use other CoVs that we identify …

The results will provide information whether bat-CoVs could use known bat and human ACE2, DPP4 or other known CoV receptors to enter cells, and allow us to determine critical receptor binding sites, viral host range, and to better predict the capacity of our CoVs to infect people. [Emphasis in original]

A research scientist will be hired at 12 months time per year to provide direct assistance and oversight of field activities in China; maintain equipment and logistics; and coordinate animal and human sample shipment to the labs in China and in the US.

Once we secure IRBs [Institutional Review Boards] for human sampling in Y1 [Year 1], we will hire three medical officers from China provincial CDCs [Centers for Disease Control] as consultants to work in Guangxi, Hunan, and Fujian during Y2-Y5. These medical officers will be responsible for IRB approved human sampling as well as maintaining cold chain for storage and shipping samples.

Dr. Zhengli Shi, Senior Virologist. [Redacted] per year in Y1 -Y5. Dr. Shi will oversee the coronavirus screening for all samples collected in China. She will work with the PI [Principal Investigator], Co-Investigators, and Senior/Key Personnel to analyze data and write manuscripts. She will also coordinate data and material sharing with the co-investigators.

RNA Extractions

We will be running RNA extractions for 1,000 bats per year (three sample per bat: oral, anal, and blood) in each year … Extracted RNA per animal will be pooled.

DNA Sequencing

In each year of the project, DNA sequencing will be performed on 3,200 samples at a cost of $2.91 per reaction….

Laboratory Supplies

We request support for in vitro infection experiments using pseudoviruses carrying the spike proteins (wild type or mutants) or live viruses in cell lines of different origins, binding affinity assays between the spike proteins (wild types or mutants) and different cellular receptor molecules, and humanized mouse experiments.

Testing predictions of CoV inter-species transmission. The following experiments will be undertaken in Year 2:

• Humanized mice with human ACE2 receptors will be infected with WIV1 and the two rescued chimeric SARS-like coronaviruses to determine the tissue tropism and pathogenicity of bat SL-CoV.
• Isolation of novel bat coronaviruses. Live virus or pseudovirus will be used to infect cells of different origin or expressing different receptor molecules. Spillover potential for each isolated virus will be assessed.
• An infectious clone of full-length MERS-CoV [Middle East Respiratory Syndrome coronavirus] will be constructed using reverse genetic method. Using the S [spike] sequence of different MERS-related viruses identified from Chinese bats, the chimeric viruses with S gene of bat MERS-related coronaviruses and backbone of the infectious clone of MERS-CoV will be constructed to study the receptor usage and infectivity of bat MERS-related coronaviruses.

• The infectious clone of WIV1 was successfully constructed using reverse genetic methods;
• Two chimeric bat SARS-like coronavirus strains were constructed by replacing the S [spike] gene in the backbone of WIV1;
• Permission to import mice with human ACE2 to China was obtained, so as to conduct the experimental infections proposed in our R01 specific aims.

In Year 3, we successfully isolated Rs4874 from the single [bat] fecal sample. Using the reverse genetic system we previously developed, we constructed two chimeric viruses with the WIV1 backbone replaced with the S [spike] gene of Rs7327 and Rs4231, respectively. Vero E6 cells were respectively infected with Rs4874, WIV1-Rs4231S and WIV1-Rs7327S, and efficient virus replication was detected by immunofluorescence assay in all infections. To assess the usage of human ACE2 by the three novel SL-CoVs, we conducted virus infectivity studies using HeLa cells with or without the expression of human ACE2. All viruses replicated efficiently in the human ACE2-expressing cells.

Using the reverse genetic methods we previously developed, infectious clones with the WIV1 [bat SARS-like coronavirus] backbone and the spike protein of SHC014, W IV16 and Rs4231, respectively, were constructed and recombinant viruses were successfully rescued. In Year 4, we performed preliminary in vivo infection of SARSr-CoVs on transgenic mice that express hACE2. Mice were infected with 105 pfu of full-length recombinant virus of W IV1 (rWIV1) and the three chimeric viruses with different spikes. Pathogenesis of the 4 SARSr-CoVs was then determined in a 2-week course. Mice challenged with rWIV1-SHC014S have experienced about 20% body weight loss by the 6th day post infection, while rWIV1 and rWIV-4231 S produced less body weight loss. In the mice infected with rWIV1 -WIV16S, no body weight loss was observed (Fig. 35a). 2 and 4 days post infection, the viral load in lung tissues of mice challenged with rWIV1-SHC014S, rWIV1-WIV16S and rWIV1-Rs4231 S reached more than 106 genome copies/g and were significantly higher than that in rWIV1-infected mice (Fig. 35b). These results demonstrate varying pathogenicity of SARSr-CoVs with different spike proteins in humanized mice.

Importantly, we are not proposing to genetically manipulate SARS-CoV over the course of this proposal. [Emphasis in original] However, we are proposing to genetically manipulate the full length bat SARSr-CoV WIV1 strain molecular clone during the course of this proposal, which is not a select agent, has not been shown to cause human infections, and has not been shown to be transmissible between humans.

All staff rosters, phone lists, or similar records depicting all employees hired by or detailed to the office of Special Counsel Jack Smith.

Exemption 6 pertains to information the release of which would constitute a clearly unwarranted invasion of personal privacy. Exemption 7(A) pertains to records or information compiled for law enforcement purposes, the release of which could reasonably be expected to interfere with enforcement proceedings.

The Federal Bureau of Investigation (FBI) misled and stonewalled Judicial Watch on a legitimate public record request involving an illegal Chinese police station opened in New York by agents of China’s communist government. The outpost is the first in the United States operated on behalf of the Fuzhou branch of the Ministry of Public Security (MPS) of the People’s Republic of China (PRC) and was surreptitiously established in an office building in Manhattan’s Chinatown to intimidate Chinese dissidents living in this country.

Back in October 2022 Judicial Watch filed a Freedom of Information Act (FOIA) request with the FBI for records concerning the foreign police station which was operated by two Chinese men who live in New York, federal authorities recently confirmed. The men, 61-year-old Harry Lu Jianwang of the Bronx and 59-year-old Chen Jinping of Manhattan, were arrested this week and charged with conspiring to act as agents of the PRC government as well as obstructing justice by destroying evidence of their communications with an MPS official. “The PRC, through its repressive security apparatus, established a secret physical presence in New York City to monitor and intimidate dissidents and those critical of its government,” said Assistant Attorney General Matthew G. Olsen of the Justice Department’s National Security Division. An assistant director of the FBI’s counterintelligence division also said this week that the case serves as a powerful reminder that China will stop at nothing to bend people to its will.

It was not the FBI’s first public acknowledgement of the illicit Chinese police station in the Big Apple. In mid-November of last year FBI Director Christopher Wray told a congressional committee that his agency was investigating an unauthorized police station run by China in New York as part of a chain in major cities around the world. A Spanish-based human rights group had just disclosed that China has dozens of overseas police service stations globally and lawmakers asked Wray about it during a Senate Homeland Security Committee hearing. The FBI director affirmed he was aware of the existence of the stations and that it is outrageous to think the Chinese police would attempt to set up shop in New York. He also said the Chinese station “violates sovereignty and circumvents standard judicial and law enforcement cooperation processes.”

The FBI has since recognized that it conducted a search of New York’s Chinese police station in October 2022 and that agents interviewed the defendants and seized their phones. “In reviewing the contents of these phones, FBI agents observed that communications between Lu and Chen, on the one hand, and the MPS Official, on the other, appeared to have been deleted,” according to a statement issued by the Department of Justice (DOJ). “In subsequent consensual interviews, Lu and Chen admitted to the FBI that they had deleted their communications with the MPS Official after learning about the ongoing FBI investigation, thus preventing the FBI from learning the full extent of the MPS’s directions for the overseas police station.” The feds knew that Lu had a longstanding relationship of trust with Chinese law enforcement and that he was tasked with carrying various activities including to assist the PRC government’s repressive activities in the U.S.

Nevertheless, the FBI wrongfully rejected Judicial Watch’s detailed public record request, denying the existence of any information involving the Chinese police station in Manhattan. In a November 18, 2022 response to Judicial Watch’s October filing the agency outright claims it has no records. “Based on the information you provided, we conducted a main and reference entity record search of the Central Records System (CRS) per our standard search policy,” the FBI writes in its response. “However, we were unable to identify records subject to the FOIPA that are responsive to your request. Therefore, your request is being closed.” The document is signed by Michael G. Seidel, who is identified as the section chief of the Record/Information Dissemination Section of the Information Management Division.

Like most government agencies the FBI is rarely forthcoming with records, which forces Judicial Watch to sue in federal court for information that should be available to the public under the law. The FBI even took advantage of the pandemic, using it as an excuse to withhold information by shutting down its electronic public records operations during a time when mandatory social distancing forced Americans and federal government employees to telework.