LPS-Responsive CHS1/Beige-Like Anchor Gene and Therapeutic Applications Thereof

Abstract

The present invention relates to a novel LPS-responsive and Beige-like Anchor gene (lrba), variants of the lrba gene, fragments of the lrba gene, and polypeptides encoded thereby. The subject invention also pertains to lrba interfering RNA, and uses thereof. In another aspect, the present invention also includes methods of inhibiting tumor growth in a patient by suppressing lrba function.

1 . A method for inhibiting the growth of a tumor in a patient, comprising administering an agent to the patient, wherein the agent suppresses LPS-responsive CHS1/beige-like anchor gene (irba) function in the patient. 
     
     
         2 . The method according to  claim 1 , wherein the agent is a polynucleotide fragment of an lrba gene, or a variant thereof, or a polypeptide fragment of an lrba gene product, or a variant thereof. 
     
     
         3 . The method according to  claim 1 , wherein the agent is a polynucleotide sequence encoding a fragment of the amino acid sequence of SEQ ID NO: 3 or SEQ ID NO: 8, and wherein the amino acid sequence is at least five amino acids in length. 
     
     
         4 . The method according to  claim 1 , wherein the agent is a polynucleotide sequence encoding an lrba BEACH domain of SEQ ID NO: 3 or SEQ ID NO: 8, or a polynucleotide sequence encoding an lrba WD domain of SEQ ID NO: 3 or SEQ ID NO: 8, or a polynucleotide encoding both an lrba BEACH domain and an lrba WD domain of SEQ ID NO: 3 or SEQ ID NO: 8. 
     
     
         5 . The method according to  claim 1 , wherein the agent is a polynucleotide encoding an amino acid sequence selected from the group consisting of:
 residues 2028 to 2071 of SEQ ID NO: 3, or a fragment thereof;   residues 2072 to 2121 of SEQ ID NO: 3, or a fragment thereof;   residues 2122 to 2162 of SEQ ID NO: 3, or a fragment thereof;   residues 2204 to 2482 of SEQ ID NO: 3, or a fragment thereof;   residues 2584 to 2628 of SEQ ID NO: 3, or a fragment thereof;   residues 2629 to 2687 of SEQ ID NO: 3, or a fragment thereof;   residues 2688 to 2769 of SEQ ID NO: 3, or a fragment thereof;   residues 2770 to 2811 of SEQ ID NO: 3, or a fragment thereof;   residues 2812 to 2856 of SEQ ID NO: 3, or a fragment thereof;   residues 2035 to 2169 of SEQ ID NO: 8, or a fragment thereof;   residues 2212 to 2489 of SEQ ID NO: 8, or a fragment thereof;   residues 2592 to 2635 of SEQ ID NO: 8, or a fragment thereof;   residues 2636 to 2694 of SEQ ID NO: 8, or a fragment thereof;   residues 2695 to 2778 of SEQ ID NO: 8, or a fragment thereof;   residues 2779 to 2819 of SEQ ID NO: 8, or a fragment thereof; and   residues 2820 to 2863 of SEQ ID NO: 8, or a fragment thereof   
     
     
         6 . The method according to  claim 3 , wherein the polynucleotide sequence is expressed within a tumor cell of the patient. 
     
     
         7 . The method according to  claim 3 , wherein the polynucleotide is contained within a vector, and wherein the vector further comprises a promoter sequence operably linked to the polynucleotide. 
     
     
         8 . The method according to  claim 3 , wherein the polynucleotide is contained within a recombinant cell, wherein said recombinant cell further comprises a promoter sequence operably linked to the polynucleotide. 
     
     
         9 . The method according to  claim 1 , wherein the agent is an antisense oligonucleotide. 
     
     
         10 . The method according to  claim 1 , wherein the agent is administered to a tumor site on the patient. 
     
     
         11 . The method according to  claim 1 , wherein the patient is a mammal. 
     
     
         12 . The method according to  claim 1 , wherein the patient is a human. 
     
     
         13 . The method according to  claim 1 , wherein the patient is suffering from a tumor of a cancer type selected from the group consisting of breast, prostate, melanoma, chronic myelogenous leukemia, cervical cancer, adenocarcinoma, lymphoblastic leukemia, colorectal cancer, and lung carcinoma. 
     
     
         14 . An isolated polynucleotide encoding an amino acid sequence selected from the group consisting of SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, and SEQ ID NO: 24, or a fragment or variant thereof, or an isolated polynucleotide that is complementary to any of the foregoing polynucleotides. 
     
     
         15 . The polynucleotide of  claim 14 , wherein said polynucleotide encodes an amino acid sequence selected from the group consisting of:
 residues 2028 to 2071 of SEQ ID NO: 3, or a fragment thereof;   residues 2072 to 2121 of SEQ ID NO: 3, or a fragment thereof;   residues 2122 to 2162 of SEQ ID NO. 3, or a fragment thereof;   residues 2204 to 2482 of SEQ ID NO: 3, or a fragment thereof;   residues 2584 to 2628 of SEQ ID NO: 3, or a fragment thereof;   residues 2629 to 2687 of SEQ ID NO: 3, or a fragment thereof;   residues 2688 to 2769 of SEQ ID NO: 3, or a fragment thereof;   residues 2770 to 2811 of SEQ ID NO: 3, or a fragment thereof;   residues 2812 to 2856 of SEQ ID NO: 3, or a fragment thereof;   residues 2035 to 2169 of SEQ ID NO: 8, or a fragment thereof;   residues 2212 to 2489 of SEQ ID NO: 8, or a fragment thereof;   residues 2592 to 2635 of SEQ ID NO: 8, or a fragment thereof;   residues 2636 to 2694 of SEQ ID NO: 8, or a fragment thereof;   residues 2695 to 2778 of SEQ ID NO: 8, or a fragment thereof;   residues 2779 to 2819 of SEQ ID NO: 8, or a fragment thereof; and   residues 2820 to 2863 of SEQ ID NO: 8, or a fragment thereof   
     
     
         16 . The polynucleotide of  claim 14 , wherein said polynucleotide is an lrba gene isoform selected from the group consisting of lrba-α, lrba-β, lrba-ε, lrba-γ, and lrba-ε. 
     
     
         17 . An isolated polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, and SEQ ID NO: 24, or a fragment or variant thereof. 
     
     
         18 . The polypeptide of  claim 17 , wherein said polypeptide comprises an amino acid sequence selected from the group consisting of:
 residues 2028 to 2071 of SEQ ID NO: 3, or a fragment thereof;   residues 2072 to 2121 of SEQ ID NO: 3, or a fragment thereof;   residues 2122 to 2162 of SEQ ID NO: 3, or a fragment thereof;   residues 2204 to 2482 or SEQ ID NO: 3, or a fragment thereof;   residues 2584 to 2628 of SEQ ID NO: 3, or a fragment thereof;   residues 2629 to 2687 of SEQ ID NO: 3, or a fragment thereof;   residues 2688 to 2769 of SEQ ID NO: 3, or a fragment thereof;   residues 2770 to 2811 of SEQ ID NO: 3, or a fragment thereof;   residues 2812 to 2856 of SEQ ID NO: 3, or a fragment thereof;   residues 2035 to 2169 of SEQ ID NO: 8, or a fragment thereof;   residues 2212 to 2489 of SEQ ID NO: 8, or a fragment thereof;   residues 2592 to 2635 of SEQ ID NO: 8, or a fragment thereof;   residues 2636 to 2694 of SEQ ID NO: 8, or a fragment thereof;   residues 2695 to 2778 of SEQ ID NO: 8, or a fragment thereof;   residues 2779 to 2819 of SEQ ID NO: 8, or a fragment thereof; and   residues 2820 to 2863 of SEQ ID NO: 8, or a fragment thereof.   
     
     
         19 . The polypeptide of  claim 17 , wherein said polypeptide is encoded by an lrba gene isoform selected from the group consisting of lrba-α, lrba-β, lrba-δ, lrba-γ, and lrba-ε. 
     
     
         20 . A recombinant host transformed with a polynucleotide encoding an amino acid sequence selected from the group consisting of SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, and SEQ ID NO: 24, or a fragment or variant thereof, wherein said polynucleotide is operatively linked to a promoter sequence. 
     
     
         21 . The recombinant host of  claim 20 , wherein said polynucleotide encodes an amino acid sequence selected from the group consisting of:
 residues 2028 to 2071 of SEQ ID NO: 3, or a fragment thereof;   residues 2072 to 2121 of SEQ ID NO: 3, or a fragment thereof;   residues 2122 to 2162 of SEQ ID NO: 3, or a fragment thereof;   residues 2204 to 2482 of SEQ ID NO: 3, or a fragment thereof;   residues 2584 to 2628 of SEQ ID NO: 3, or a fragment thereof;   residues 2629 to 2687 of SEQ ID NO: 3, or a fragment thereof;   residues 2688 to 2769 of SEQ ID NO: 3, or a fragment thereof;   residues 2770 to 2811 of SEQ ID NO: 3, or a fragment thereof;   residues 2812 to 2856 of SEQ ID NO: 3. or a fragment thereof;   residues 2035 to 2169 of SEQ ID NO: 8, or a fragment thereof;   residues 2212 to 2489 of SEQ ID NO: 8, or a fragment thereof;   residues 2592 to 2635 of SEQ ID NO: 8, or a fragment thereof;   residues 2636 to 2694 of SEQ ID NO: 8, or a fragment thereof;   residues 2695 to 2778 of SEQ ID NO: 8, or a fragment thereof;   residues 2779 to 2819 of SEQ ID NO: 8, or a fragment thereof; and   residues 2820 to 2863 of SEQ ID NO: 8, or a fragment thereof.   
     
     
         22 . The recombinant host of  claim 20 , wherein said host is a prokaryotic cell. 
     
     
         23 . The recombinant host of  claim 20 , wherein said host is a eukaryotic cell. 
     
     
         24 . A vector comprising a polynucleotide encoding an amino acid sequence selected from the group consisting of SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6. SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, and SEQ ID NO: 24, or a fragment or variant thereof. 
     
     
         25 . The vector of  claim 24 , wherein said vector is an adenovirus. 
     
     
         26 . The vector of  claim 24 , wherein said polynucleotide encodes an amino acid sequence selected from the group consisting of:
 residues 2028 to 2071 of SEQ ID NO: 3, or a fragment thereof;   residues 2072 to 2121 of SEQ ID NO: 3, or a fragment thereof;   residues 2122 to 2162 of SEQ ID NO: 3, or a fragment thereof;   residues 2204 to 2482 of SEQ ID NO: 3, or a fragment thereof;   residues 2584 to 2628 of SEQ ID NO: 3, or a fragment thereof;   residues 2629 to 2687 of SEQ ID NO: 3, or a fragment thereof;   residues 2688 to 2769 of SEQ ID NO: 3. or a fragment thereof;   residues 2770 to 2811 of SEQ ID NO: 3, or a fragment thereof;   residues 2812 to 2856 of SEQ ID NO: 3, or a fragment thereof;   residues 2035 to 2169 of SEQ ID NO: 8, or a fragment thereof;   residues 2212 to 2489 of SEQ ID NO: 8, or a fragment thereof;   residues 2592 to 2635 of SEQ ID NO: 8, or a fragment thereof;   residues 2636 to 2694 of SEQ ID NO: 8, or a fragment thereof;   residues 2695 to 2778 of SEQ ID NO: 8, or a fragment thereof;   residues 2779 to 2819 of SEQ ID NO: 8, or a fragment thereof and   residues 2820 to 2863 of SEQ ID NO: 8, or a fragment thereof.   
     
     
         27 . The vector of  claim 24 , wherein said vector further comprises a promoter sequence operatively linked to said polynucleotide. 
     
     
         28 . An interfering RNA molecule that suppresses mammalian LPS-responsive CHS1/beige-like anchor gene (lrba) function. 
     
     
         29 . The method according to  claim 1 , wherein the agent is selected from the group consisting of a dominant negative mutant, an interfering RNA molecule, and a peptidomimetic. 
     
     
         30 . The method according to  claim 29 , wherein the dominant negative mutant is a polynucleotide encoding the BEACH domain of SEQ ID NO: 3 or SEQ ID NO: 8, a polynucleotide encoding a WD domain of SEQ ID NO: 3 or SEQ ID NO: 8, or a polynucleotide encoding both the BEACH domain of SEQ ID NO: 3 or SEQ ID NO: 8 and a WD domain of SEQ ID NO: 3 or SEQ ID NO: 8. 
     
     
         31 . A method of screening an individual for cancer comprising obtaining a biological sample from the individual and determining the amount of lrba gene expression in the biological sample. 
     
     
         32 . The method according to claim  34 , further comprising comparing the determined amount of lrba gene expression in the sample with that found in a normal population.