Peripheral-type benzodiazepine receptor: a tool for detection, diagnosis, prognosis, and treatment of cancer
AssigneeGeorgetown University
InventorsVASSILIOS PAPADOPOULOS, MARTINE CULTY
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Abstract
The expression and subcellular localization of peripheral-type benzodiazepine receptors (PBR) is shown in this application to correlate with the metastatic potential of cells, and increased cell proliferation. Inhibition of PBR expression, function or stability results in a decrease in cell proliferation. Compositions and methods for regulating and/or monitoring PBR and its expression are useful for the detection, diagnosis, prognosis and treatment of solid tumors, in particular, breast cancer.
Claims36
A composition for decreasing cell proliferation, comprising an antibody, drug or agent which reduces or inhibits peripheral-type benzodiazepine receptors (PBR) function.
A method for determining aggressive phenotype of a tumor comprising:
- (i) contacting a tumor cell with antibodies which recognize PBR;
- and
- (ii) detecting the presence of absence of a complex formed between PBR and antibodies specific for PBR in the nucleus of said tumor cells wherein the presence of a complex in the nucleus indicates the presence of aggressive tumor phenotype.
A method for determining aggressive tumor phenotype comprising:
- (i) contacting a tumor cell with antibodies which recognize PBR;
- (ii) detecting the presence of absence of a complex formed between PBR and antibodies specific for PBR in the nucleus of said tumor cells;
- and
- (iii) comparing the amount of complex formed with an amount of PBR complexed from a normal cell, wherein in increase in complex over normal indicates the presence of an aggressive tumor phenotype.
A method for diagnosing an aggressive tumor phenotype comprising:
- (i) contacting a tumor tissue sample with oligonucleotides which recognize PBR RNA;
- (ii) detecting the presence or absence of a duplex formed between PBR RNA in said sample and oligonucleotides specific therefor;
- (iii) and comparing it to the amount of duplex formed in a normal tissue sample, wherein an increase in duplex in the suspected tissue over normal indicates the presence of an aggressive tumor phenotype.
A therapeutic method for the treatment or amelioration of diseases and processes that are mediated by increased cell proliferation comprising the steps of administering to an individual in need of such treatment antibodies, drugs or agents which reduce or eliminate the function of PBR in a pharmaceutically acceptable diluent in a pharmaceutically acceptable amount.
A composition for reducing cancer growth comprising an inhibitor of PBR selected from the group comprising
- (i) PBR specific antibody;
- (ii) PBR antagonist;
- (iii) PBR antisense oligonucleotide;
- and
- (iv) a natural or synthetic inhibitor of PBR.
A therapeutic method for the treatment or amelioration of diseases and processes that are mediated by reduced cell proliferation comprising the step of increasing PBR by administering PBR in a pharmaceutically acceptable diluent in a pharmaceutically acceptable amount such that cell proliferation is increased.
An in vitro method for testing possible agents or drugs for cell proliferation inhibitory activity said method comprising measuring ability of said agent or drug to decrease PBR activity in an in vitro assay.
An in vitro method for testing possible drugs or agent which promote cell proliferation said method comprising measuring ability of said agent or drug to increase PBR function in an in vitro assay.
A composition for detecting PBR comprising at least one of the following:
- anti PBR antibody, natural PBR ligand, and synthetic PBR ligand.
A method for detecting the level of PBR in cells using the polymerase chain reaction said method comprising:
- (i) extracting RNA from a sample;
- (ii) reverse transcribing said RNA into cDNA
- (ii) contacting said cDNA with
- (a) at least four nucleotide triphosphates,
- (b) a primer that hybridizes to PBR cDNA, and
- (c) an enzyme with polynucleotide synthetic activity,
- under conditions suitable for the hybridization and extension of said first primer by said enzyme, whereby a first DNA product is synthesized with said DNA as a template therefor, such that a duplex molecule is formed;
- (iii) denaturing said duplex to release said first DNA product from said DNA;
- (iv) contacting said first DNA product with a reaction mixture comprising:
- (a) at least four nucleotide triphosphates,
- (b) a second primer that hybridizes to said first DNA, and
- (c) an enzyme with polynucleotide synthetic activity,
- under conditions suitable for the hybridization and extension of said second primer by said enzyme, whereby a second DNA product is synthesized with said first DNA as a template therefor, such that a duplex molecule is formed;
- (v) denaturing said second DNA product from said first DNA product;
- (vi) repeating steps iii-vi for a sufficient number of times to achieve linear production of said first and second DNA products;
- (vii) fractionating said first and second DNA products generated from said PBR cDNA;
- and
- (viii) comparing the level of PBR cDNA with the level of PBR cDNA from a normal cell;
- wherein, an increase in PBR level over normal cells indicates an aggressive tumor phenotype.
A method for determining the aggressive phenotype of a tumor cell detecting PBR RNA in said cell and comparing the level of PBR RNA to the level of PBR RNA from a normal cell wherein an increase over normal in PBR RNA in the tumor cell indicates an aggressive tumor phenotype.
A cDNA comprising the polynucleotide sequence specified in SEQ ID NO:1 or SEQ ID NO:2 encoding a polypeptide comprising the sequence specified in SEQ ID NO:3.
A PBR negative cell comprising an inactivated PBR gene.
Dependencies are inferred from the claim text.