Alnylam and Collaborators Publish Novel Method for Detection of Tissue-Specific, RNAi-Mediated Gene Silencing

  Alnylam and Collaborators Publish Novel Method for Detection of
  Tissue-Specific, RNAi-Mediated Gene Silencing

     – New Results Published in the Journal “RNA” Demonstrate Utility of
     Circulating Extracellular RNA Detection (cERD) Method in Quantifying
 Circulating Messenger RNA and Micro-RNA in Serum and Cerebrospinal Fluid to
Monitor Tissue-Specific RNA Knockdown and to Confirm RNAi Proof of Mechanism –

Business Wire

CAMBRIDGE, Mass. -- December 20, 2013

Alnylam Pharmaceuticals, Inc. (Nasdaq: ALNY), a leading RNAi therapeutics
company, announced today the publication of a new paper describing a novel
technique called circulating Extracellular RNA Detection (cERD). The paper,
titled “Tissue-specific gene silencing monitored in circulating RNA” (Sehgal
et al., RNA, doi:10.1261/rna.042507.113) appears online as an Advance Article
in the journal RNA. The cERD method enables the quantification of circulating
messenger RNA (mRNA) and micro-RNA (miRNA) as a way of monitoring
tissue-specific RNA silencing. It also enables confirmation of an
RNAi-mediated mechanism of action using a PCR-based technique. The cERD method
could have broad applicability in clinical studies since it allows for
monitoring of tissue-specific mRNA levels using a non-invasive technique.

“Detection of target gene knockdown is typically made possible by measurement
and quantification of mRNA levels in tissue samples and, if the target protein
is secreted, corresponding protein levels in serum or plasma. Further,
establishing proof of mechanism for RNAi therapeutics involves analysis of
RNAi-mediated target mRNA cleavage with a PCR-based technique in tissue
samples. However, the utility of these techniques is limited, since many
proteins do not enter circulation, and the need to collect biopsies creates a
significant challenge to the routine monitoring of mRNA expression levels or
the RNAi mechanism of action in tissues,” said Rachel Meyers, Ph.D., Vice
President of Research and RNAi Lead Development at Alnylam. “These new
published data establish the applicability of the cERD method in multiple
species, including rats and non-human primates, and thus the general utility
of measuring circulating serum mRNA to confirm RNAi-mediated tissue gene
silencing. Specifically, we have shown that changes in circulating RNA levels
correspond closely to changes in tissue RNA levels, suggesting that RNA levels
from biological fluids provide an accurate ‘real-time’ representation of
tissue RNA status. These findings are important to the development of RNAi
therapeutics, as the cERD method could be extended to the clinical setting to
allow the routine, accurate, and frequent measurement of organ-specific target
gene knockdown and/or RNAi mechanism without the need for tissue biopsies.”

The new research published in RNA shows that mRNAs encoding tissue-specific
gene transcripts can be detected in biological fluids. Specifically,
RNAi-mediated target mRNA silencing in the liver by systemic siRNA
administration resulted in quantitative reductions in serum mRNA levels that
were shown to closely correlate with the degree and kinetics of mRNA silencing
in liver tissue. Further, by identifying the predicted RNAi cleavage products
via analysis of circulating RNA in serum, the authors demonstrated the use of
cERD to verify the RNAi mechanism for target gene knockdown. For example,
Alnylam recently published results in which the RNAi mechanism was confirmed
using the cERD method in serum collected from humans participating in a Phase
I clinical trial (Coelho et al., N Engl J Med 2013;369:819-29). In addition,
administration of an anti-miR directed against a liver-specific miRNA led to
decreased levels of the miRNA in serum, showing that cERD can also be applied
to monitoring pharmacodynamics for microRNA therapeutics. Moreover, the
authors quantified expression of an adenoviral transgene in the liver by
measuring levels of circulating mRNA, suggesting that measurement of mRNA
levels in serum can be applied to the analysis of exogenously delivered genes
and thus can be a useful tool for gene therapy applications. Finally, using
the cERD method, the authors monitored and quantified the degree of silencing
of a brain-expressed mRNA in cerebrospinal fluid following intraparenchymal
siRNA infusion to the central nervous system.

About RNA Interference (RNAi)

RNAi (RNA interference) is a revolution in biology, representing a
breakthrough in understanding how genes are turned on and off in cells, and a
completely new approach to drug discovery and development. Its discovery has
been heralded as “a major scientific breakthrough that happens once every
decade or so,” and represents one of the most promising and rapidly advancing
frontiers in biology and drug discovery today which was awarded the 2006 Nobel
Prize for Physiology or Medicine. RNAi is a natural process of gene silencing
that occurs in organisms ranging from plants to mammals. By harnessing the
natural biological process of RNAi occurring in our cells, the creation of a
major new class of medicines, known as RNAi therapeutics, is on the horizon.
Small interfering RNA (siRNA), the molecules that mediate RNAi and comprise
Alnylam’s RNAi therapeutic platform, target the cause of diseases by potently
silencing specific mRNAs, thereby preventing disease-causing proteins from
being made. RNAi therapeutics have the potential to treat disease and help
patients in a fundamentally new way.

About Alnylam Pharmaceuticals

Alnylam is a biopharmaceutical company developing novel therapeutics based on
RNA interference, or RNAi. The company is leading the translation of RNAi as a
new class of innovative medicines with a core focus on RNAi therapeutics
toward genetically defined targets for the treatment of serious,
life-threatening diseases with limited treatment options for patients and
their caregivers. These include: patisiran (ALN-TTR02), an intravenously
delivered RNAi therapeutic targeting transthyretin (TTR) for the treatment of
TTR-mediated amyloidosis (ATTR) in patients with familial amyloidotic
polyneuropathy (FAP); ALN-TTRsc, a subcutaneously delivered RNAi therapeutic
targeting TTR for the treatment of ATTR in patients with familial amyloidotic
cardiomyopathy (FAC); ALN-AT3, an RNAi therapeutic targeting antithrombin (AT)
for the treatment of hemophilia and rare bleeding disorders (RBD); ALN-AS1, an
RNAi therapeutic targeting aminolevulinate synthase-1 (ALAS-1) for the
treatment of porphyria including acute intermittent porphyria (AIP); ALN-CC5,
an RNAi therapeutic targeting complement component C5 for the treatment of
complement-mediated diseases; ALN-PCS, an RNAi therapeutic targeting PCSK9 for
the treatment of hypercholesterolemia; ALN-TMP, an RNAi therapeutic targeting
TMPRSS6 for the treatment of beta-thalassemia and iron-overload disorders;
ALN-AAT, an RNAi therapeutic targeting alpha-1-antitrypsin (AAT) for the
treatment of AAT deficiency liver disease; and ALN-ANG, an RNAi therapeutic
for the treatment of genetic forms of mixed hyperlipidemia and severe
hypertriglyceridemia, amongst other programs. As part of its “Alnylam 5x15”
strategy, the company expects to have five RNAi therapeutic products for
genetically defined diseases in clinical development, including programs in
advanced stages, on its own or with a partner by the end of 2015. Alnylam has
additional partnered programs in clinical or development stages, including
ALN-RSV01 for the treatment of respiratory syncytial virus (RSV) infection and
ALN-VSP for the treatment of liver cancers. The company’s leadership position
on RNAi therapeutics and intellectual property have enabled it to form major
alliances with leading companies including Merck, Medtronic, Novartis, Biogen
Idec, Roche, Takeda, Kyowa Hakko Kirin, Cubist, Ascletis, Monsanto, Genzyme,
and The Medicines Company. In addition, Alnylam holds an equity position in
Regulus Therapeutics Inc., a company focused on discovery, development, and
commercialization of microRNA therapeutics. Alnylam has also formed Alnylam
Biotherapeutics, a division of the company focused on the development of RNAi
technologies for applications in biologics manufacturing, including
recombinant proteins and monoclonal antibodies. Alnylam’s VaxiRNA™ platform
applies RNAi technology to improve the manufacturing processes for vaccines;
GlaxoSmithKline is a collaborator in this effort. Alnylam scientists and
collaborators have published their research on RNAi therapeutics in over 100
peer-reviewed papers, including many in the world’s top scientific journals
such as Nature, Nature Medicine, Nature Biotechnology, Cell, the New England
Journal of Medicine, and The Lancet. Founded in 2002, Alnylam maintains
headquarters in Cambridge, Massachusetts. For more information, please visit

Alnylam Forward-Looking Statements

Various statements in this press release concerning Alnylam’s future
expectations, plans and prospects, including without limitation, Alnylam’s
expectations regarding its “Alnylam 5x15” product strategy, Alnylam’s views
with respect to the potential for RNAi therapeutics, and Alnylam’s views with
respect to the use of the cERD method in clinical studies constitute
forward-looking statements for the purposes of the safe harbor provisions
under The Private Securities Litigation Reform Act of 1995. Actual results may
differ materially from those indicated by these forward-looking statements as
a result of various important factors, including, without limitation,
Alnylam’s ability to discover and develop novel drug candidates and delivery
approaches, successfully demonstrate the efficacy and safety of its drug
candidates, the pre-clinical and clinical results for its product candidates,
which may not support further development of product candidates, actions of
regulatory agencies, which may affect the initiation, timing and progress of
clinical trials, obtaining, maintaining and protecting intellectual property,
Alnylam’s ability to enforce its patents against infringers and defend its
patent portfolio against challenges from third parties, obtaining regulatory
approval for products, competition from others using technology similar to
Alnylam’s and others developing products for similar uses, Alnylam’s ability
to obtain additional funding to support its business activities and establish
and maintain strategic business alliances and new business initiatives,
Alnylam’s dependence on third parties for development, manufacture, marketing,
sales and distribution of products, the outcome of litigation, and unexpected
expenditures, as well as those risks more fully discussed in the “Risk
Factors” filed with Alnylam’s most recent Quarterly Report on Form 10-Q filed
with the Securities and Exchange Commission (SEC) and in other filings that
Alnylam makes with the SEC. In addition, any forward-looking statements
represent Alnylam’s views only as of today and should not be relied upon as
representing its views as of any subsequent date. Alnylam explicitly disclaims
any obligation to update any forward-looking statements.


Alnylam Pharmaceuticals, Inc.
Cynthia Clayton, 617-551-8207
Vice President, Investor Relations and Corporate Communications
Amanda Sellers (Media), 202-955-6222 x2597
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