[Federal Register Volume 64, Number 152 (Monday, August 9, 1999)]
[Notices]
[Pages 43200-43201]
From the Federal Register Online via the Government Publishing Office [www.gpo.gov]
[FR Doc No: 99-20455]
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DEPARTMENT OF HEALTH AND HUMAN SERVICES
National Institutes of Health
Government-Owned Invention; Availability for Licensing: ``Novel
Method and Composition to Induce Apoptosis in Tumor Cells''
AGENCY: National Institutes of Health, Public Health Service, DHHS.
ACTION: Notice
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SUMMARY: The invention listed below is owned by an agency of the U.S.
Government and is available for licensing in the U.S. in accordance
with 35 U.S.C. 207 to achieve expeditious commercialization of results
of federally funded research and development.
ADDRESSES: Licensing information and a copy of the U.S. patent
application referenced below may be obtained by contacting J. R. Dixon,
Ph.D., at the Office of Technology Transfer, National Institutes of
Health, 6011 Executive Boulevard, Suite 325, Rockville, Maryland 20852-
3804 (telephone 301/496-7056 ext 206; fax 301/402-0220; E-Mail:
[email protected]). A signed Confidential Disclosure Agreement is required
to receive a copy of any patent application.
SUPPLEMENTARY INFORMATION: Invention Title: ``Apoptosis Inducing Agents
and Methods'' Inventors: Drs. Lucio Miele (U.S.F.D.A.) and Leslie L.
Shelly (NICHD) USPA SN: 60/102,816 [=DHHS Ref. No. E-176-98/0]--Filed
with the U.S.P.T.O. October 2, 1998.
Apoptosis or programmed cell death is caused by many anti-tumor
drugs and by radiation therapy. These treatment modalities cause
apoptosis in tumor cells and in many normal cells in the body. As
cancer cells progress towards more aggressive forms, they often become
highly resistant to drug- or radiation-induced apoptosis, generally
through the loss of function p53, a gene which can trigger apoptosis in
response to DNA damage. Thus, novel strategies to induce apoptosis in
tumor cells, especially p53-deficient cells, is an attractive and an
active area of research.
An antisense molecule is a DNA or RNA which has the opposite
beginning to end orientation compared to the ``normal'' gene. These
molecules reduce the expression of the target gene by forming pairs
with its ``normal'' DNA and RNA. Notch-1 is a gene which is known to be
important in controlling cell differentiation in many organisms. Notch-
1 is expressed at high levels in several human tumors. However, its
function in tumor cells has not been characterized. So far, its role in
maintaining tumor cell survival has not been identified. Using a model
constituted by a p53-deficient mouse leukemia cell line, NIH scientists
found that: 1.) Antisense synthetic DNA oligonucleotides and stable
incorporation of an antisense gene (a model for gene therapy) targeting
notch-1, when given together with a differentiation-inducing antitumor
drug, cause the cells to respond by massive apoptosis rather than
differentiation; 2.) stable incorporation of an antisense notch-1 gene
increases apoptosis in these cells even in the absence of any antitumor
drugs. This suggests that antisense notch-1 treatment, by antisense
oligonucleotides or by gene therapy, may be used alone or together with
anti-cancer drugs to cause apoptosis in tumor cells.
[[Page 43201]]
The notch gene belongs to a family of epidermal growth factor
(``EGF'') like homeotic genes, which encode transmembrane proteins with
a variable number of cystgeine-rich EGF-like repeats in the
extracellular region. Four notch genes have been described in mammals,
which include notch-1, notch-2, notch-3, and notch-4 (Int-3), which
have been implicated in the differentiation of the nervous system and
other structures. The EGF-like proteins Delta and Serrate have been
identified as ligands of notch-1.
Mature notch proteins are heterodimeric receptors derived from the
cleavage of notch pre-proteins into an extracellular subunit
(NEC) containing multiple EGF-Like repeats and a
transmembrane subunit including intracellular region (Ntm).
Notch activation results from the binding of ligands expressed by
neighboring cells, and signaling from activated notch involves a
network of transcription regulators.
Alteration of notch-1 signaling or expression may contribute to
tumorigenesis. Deletions of the extracellular portion of human notch-1
are associated with about 10% of the cases of T-Cell acute
lymphoblastic leukemia. Truncated forms of notch-1 cause T-Cell
lymphomas when introduced into mouse bone marrow stem cells and are
onogenic in rat kidney cells. The human notch-1 gene is in a
chromosomal region (9q34) associated with hematopoietic malignancies of
lymphoid, myeloid, and erythroid lineage. Additionally, strikingly
increased expression of notch-1 has been documented in a number of
human tumors including cervical cancer, colon tumors, lung tumors, and
pre-neoplastic lesions of the uterine cervix.
Notch antisense oligonucleotides (or other molecules that interfere
with the expression or function of notch) could be therapeutically
administered to treat or prevent tumors. It has not been found that
administration of notch antisense oligonucleotide alone is ineffective
as an anti-neoplastic treatment. The present invention has overcome
this problem by combining the administration of a cell differentiation
agent with a molecule that interferes with the expression or function
of a notch protein (such as the notch-1 protein). This combination of
approaches has unexpectedly been found to induce apoptosis in
neoplastic cells, and provide a useful therapeutic application of this
technology. The method of the present invention includes inducing
apoptosis in a target cell by inhibiting a cell fate determining
function of a notch protein in the target cell at a time when the cell
is undergoing differentiation. In particular, the target cell is
induced to differentiate and upregulate notch expression, so that
interference with notch expression or function causes the target cell
to commit to an apoptotic pathway. Inhibition of notch expression or
interference with its function can include exposing the cell to a notch
protein antisense oligonucleotide that includes at least six
nucleotides that comprise a sequence complementary to at least a
portion of the RNA transcript of a notch gene (such as the notch-1
gene), and is hybridizable to the RNA transcript. Although the
antisense oligonucleotide can be hybridizable to any region of the RNA
transcript, particular oligonucleotides that have been found to be
useful are antisense oligonucleotides to the notch-1 EGF repeat region,
Lin/notch region, or ankyrin region. Alternatively the molecule can be
a monoclonal antibody that antagonizes the function of a notch protein
in the cell.
In particular the tumor cell is one that is characterized by
increased activity or increased expression of a notch protein, such as
a notch-1 or notch-2 protein. Examples of tumor types that over express
notch-1 include cervical cancer, breast cancer, colon cancer, melanoma,
seminoma, lung cancer and hematopoietic malignancies, such as erythroid
leukemia, myeloid leukemia, (such as chronic or acute myelogenous
leukemia), neuroblastoma and medulloblastoma. The differentiation
inducing agent to which the cell is exposed can be selected from a
broad variety of agents, including retinoids, polar compounds (such as
hexamethylene bisacetanmide), short chain fatty acids, organic acids,
Vitamin D derivatives, cyclooxygenase inhibitors, arachidonate
metabolism inhibitors, ceramides, diacylglycerol, cyclic nucleotide
derivatives, hormones, hormone antagonists, biologic promoters of
differentiation, and derivatives of any of these agents.
Technology
This invention provides a method and pharmaceutical composition for
treating a tumor by causing apoptosis in tumor cells that expresses
notch-1 protein, and in particular cells that exhibit increased
expression of notch-1. Hence, this technology discloses methods and
compositions to induce apoptosis in cells that over express the notch
proteins. A cell fate determining function of notch is specifically
disrupted at a time when the cell is undergoing differentiation, which
causes the cell to undergo apoptosis. The invention includes therapies
for tumors that over express a notch protein (such as notch-1) by
inducing differentiation of the cells in the tumor with a
differentiation inducing agent such as hexamethylene bisacetamide and
other such differentiation agents. At a time during which
differentiation has been promoted, and the cell is susceptible to
interference with the anti-apoptosis effect of notch, the function of
the notch protein is disrupted. Disruption of notch function can be
achieved, for example, by the expression of antisense oligonucleotides
that specifically interfere with expression of the notch protein on the
cell, or by monoclonal antibodies that specifically bind to notch and
inactivate it. This technology represents a novel method to induce
apoptosis in tumor cells.
The above mentioned invention is available, including any available
foreign intellectual property rights, for licensing.
Dated: August 3, 1999.
Jack Spiegel,
Director, Division of Technology Development and Transfer, Office of
Technology Transfer, National Institutes of Health.
[FR Doc. 99-20455 Filed 8-6-99; 8:45 am]
BILLING CODE 4140-01-P