Tuesday, October 4, 2011

Researcher Fired Amidst New Controversy

http://news.sciencemag.org/scienceinsider/2011/10/xmrv-researcher-fired.html

Chronic Fatigue Syndrome Researcher Fired Amidst New Controversy
by Jon Cohen on 4 October 2011, 6:19 PM

Judy Mikovits has had a rough few weeks. On 22 September, Science
published online a nine-lab study widely seen as the final blow to the
theory, championed by Mikovits and colleagues in an October 2009
Science paper, that a recently detected mouse retrovirus might play a
causal role in chronic fatigue syndrome (CFS). A letter in the same
issue of Science from one of the contributing labs to the 2009 report
revealed that a contamination had marred its contribution—PCR
detection and sequencing of the mouse virus, dubbed XMRV. Mikovits and
colleagues defended the validity of the rest of the study, known as
Lombardi et al., which detected the virus by several other methods, so
Science issued a rare partial retraction of the original paper.

Then on 29 September, Mikovits was fired from her job as research
director of the Whittemore Peterson Institute for Neuro-Immune Disease
(WPI), a private organization in Reno, Nevada, devoted to CFS research
and treatment. Both Mikovits and WPI's CEO, Annette Whittemore, say
the firing was not related to the XMRV theory's demise.

The very next day, a graduate student who writes a snarky blog that
has been highly critical of Mikovits and the XMRV theory raised
questions about whether a figure in Lombardi et al. had been
misrepresented. Science Executive Editor Monica Bradford said in a
statement that the journal is investigating the allegation. "As is our
policy in cases of alleged figure manipulation, we follow up with the
research authors as soon as our own review of the allegation is
complete," said Bradford. "Science takes all such matters seriously
and seeks to respond thoroughly and efficiently."

The furor revolves around an image—the bottom half of Figure 2C in
Lombardi et al.—that shows XMRV proteins in CFS patients but not
healthy controls. In her blog known as ERV (endogenous retroviruses),
Abbie Smith on 30 September noted the striking similarities between
Figure 2C and a slide Mikovits presented at a CFS meeting in Ottawa,
Canada, on 23 September. Smith, who is working on her doctoral
dissertation at the University of Oklahoma, Oklahoma City, and studies
HIV, wrote that an anonymous tipster had pointed out to her that the
two images looked identical but had different patient numbers and
experimental conditions. Smith questioned whether this was a simple
mistake or an attempt to recycle old data to make a new argument.

The Ottawa slide supported Mikovits's contention that even if XMRV
could not be detected in CFS patients, other gammaretroviruses still
lurked in their chromosomes. Mikovits described how she had treated
cells from two CFS patients with a chemical, 5-Azacytidine, that takes
methyl groups off DNA. This procedure prods cells that harbor latent
versions of retroviruses to produce them, and the image on the slide
showed the resultant proteins in what's known as a Western blot gel.
In Lombardi et al. what appears to be the same image shows "XMRV
proteins" and makes no mention of 5-Azacytidine use.

Mikovits's collaborator, Francis Ruscetti of the National Cancer
Institute (NCI) in Frederick, Maryland, who ran all of the Western
blots, confirms that the Ottawa slide uses the same image that appears
in Lombardi et al. Ruscetti and Mikovits, in a joint e-mail to Science
for this article, said many patients and their doctor, Daniel Peterson
(who since has had a falling out with WPI), knew the original coded
numbers, so the researchers changed them for the Science publication
to "protect the patient privacy." Ruscetti says it was a mistake for
Mikovits to have used the original patient codes in Ottawa. "We were
under so much pressure, we missed it," says Ruscetti.

As far as the use of 5-Azacytidine, Ruscetti and Mikovits stressed in
their e-mail that "there was no attempt in the original paper to hide
anything." They say for the purposes of Lombardi et al., the use of
5-Azacytidine was not germane: They were simply trying to demonstrate
that CFS patients had viral proteins not seen in controls. By the time
of the Ottawa meeting, they say they realized that this experiment did
not in fact show XMRV but proteins from a broader family of
gammaretroviruses.

After Lombardi et al. appeared, several laboratories had reported that
they could not detect XMRV in CFS patients. But Ruscetti and Mikovits
note that most of these studies relied on polymerase chain reaction,
which used DNA sequences of XMRV to fish out pieces of the virus from
blood samples. Those tests, they point out, would have missed other
gammaretroviruses with different sequences. The Western blot assay
they used in Lombardi et al. just so happened to cast a wider net that
uncovered proteins from any member of the gammaretrovirus family. In
addition, they say, the use of 5-Azacytidine made clear that these
infections would be missed in routine assays, as these viruses often
exist in a latent state.

On the ERV blog, Smith and others also argued that the study lacked
proper controls: the healthy controls in this experiment did not have
5-Azacytidine added to their samples. "My [principal investigator]
would say 'Why did you run this gel?' if I handed it to him," wrote
Smith.

Vinay Pathak, a retrovirologist at NCI who earlier damaged the
XMRV/CFS theory with a study in published in Science that documented
how the virus was accidentally created in laboratory experiments, says
he is "bewildered" by Ruscetti's and Mikovits's explanations about
Figure 2C. "If [5-Azacytidine] was used in the original experiment,
it's an egregious error to leave it out of the Science paper," says
Pathak. "It makes a difference how I would interpret the results."

Jonathan Stoye, a retrovirologist who once supported the XMRV/CFS
hypothesis but subsequently changed his mind after his own studies
failed to replicate the finding, says its time to retract Lombardi et
al. in its entirety. "I think there's a point where Science has to
say, there is no substance to this paper," says Stoye. "It was
published with a message, and that message is gone."

Neither Lombardi et al. nor the questions about the slides were
mentioned in Whittemore's 30 September letter formally terminating
Mikovits's contract, sent the day after the two had a heated phone
conversation. In that letter Whittemore charged her high-profile
researcher with being "insubordinate and insolent." Mikovits was
immediately locked out of her lab.

Three letters between Whittemore and Mikovits say the firing hinged on
Mikovits's failure to pass on a cell line that was sent to Vincent
Lombardi, the first author of the October 2009 Science paper who runs
UNEVX (formerly known as VIPDx), a diagnostic laboratory owned by WPI.
Until recently, the lab sold a test for XMRV and related viruses.

In a 1 October written response to Whittemore, Mikovits contended that
it was "completely appropriate" for her, as research director, not to
give Lombardi the cell line. The cell line was not related to studies
of the gammaretroviruses, but Lombardi wanted to use it for
experiments connected to a grant Mikovits had secured from the U.S.
National Institutes of Health to study possible causes of CFS.
Mikovits contended that Lombardi "was unwilling to take my direction"
and should not be undertaking a new project "while neglecting his
other duties." She also questioned his ability to carry out that
experiment.

Annette Whittemore issued a statement to Science in which she strongly
defended Lombardi's performance. "Dr. Lombardi is a valued and
important part of our team, and conducts his research work
accordingly," wrote Whittemore. "While personnel matters are generally
confidential, the statements made by Judy Mikovits are wrong, without
merit and those of a disgruntled former employee."

In an interview with Science, Mikovits contended that her firing was
also linked to a longstanding battle about WPI's decision to sell,
through VIPDx/UNEVX, a test for human gammaretroviruses. The lab began
offering the tests, which cost around $500, shortly after Lombardi et
al. reported a link between XMRV and CFS. Some patients who tested
positive went on to take antiretroviral drugs. "I said, 'No, no, no,
no,' " says Mikovits of the test. "I've asked them for the better part
of 2 years to show me that what we got in Lombardi et al. is the same
thing they're selling to patients."

The issue came to a head with the recent publication by Science of the
nine-lab study. The so-called Blood Working Group, which included the
labs run by Mikovits and Ruscetti, failed to reliably find XMRV or
other gammaretroviruses in blinded samples from people who previously
had tested positive for these viruses. Both Mikovits and Ruscetti
co-authored the paper, which invalidated their own assays for XMRV.
WPI says UNEVX has stopped offering the diagnostic tests, but did not
give details about the timing or reasons.

Whittemore, who refers to CFS as myalgic encephalomyelitis (ME)—a
common name for CFS in Europe—stressed that the institute remains
devoted to studying human gammaretroviruses in "ME and related
diseases" and that no one there "would ever put self interest ahead of
research or finding the causes of ME."

Mikovits, who says she currently does not even have access to her
laboratory notebooks, is looking for another institution to continue
her work.


With reporting by Martin Enserink

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