NOVEL mRNA SPLICE VARIANT OF THE DOUBLECORTIN-LIKE KINASE GENE AND ITS USE IN DIAGNOSIS AND THERAPY OF CANCERS OF NEUROECTODERMAL ORIGIN
The present invention relates to novel nucleic acid and protein molecules and their use in cancer therapy and diagnosis.
1 . A method of treating cancer in a subject comprising providing to a subject in need thereof an effective amount of a nucleic acid fragment of SEQ ID NO:1 or SEQ ID NO:2, or a fragment of a variant of SEQ ID NO:1 or SEQ ID NO:2, which nucleic acid fragment significantly reduces in cancer cells in the subject the amount of a doublecortin-like (DCL-protein, the sequence of which is SEQ ID NO:3 or SEQ ID NO:4, thereby treating said cancer.
2 . The method according to claim 1 , wherein said cancer is of neuroectodermal origin.
3 . The method according to claim 2 wherein said cancer is neuroblastoma, medulloblastoma, glioblastoma, oligodendroglioma, oligoastrocytoma, astrocytoma, neurofibroma, ependymoma, a malignant peripheral nerve sheath tumors, ganglioneuroma, Schwannoma, rhabdomyosarcoma, retinoblastoma, small cell lung carcinoma, adrenal pheochromocytoma, a primitive peripheral neuroectodermal tumor, Ewing's sarcoma or melanoma.
4 . The method according to claim 1 , wherein the nucleic acid fragment or variant is an antisense RNA oligonucleotide, an antisense DNA oligonucleotide or a double stranded small interfering RNA.
5 . A sense and/or antisense nucleic acid fragment of SEQ ID NO:1 or SEQ ID NO:2, or a fragment of a variant of SEQ ID NO:1 or SEQ ID NO:2, which that said nucleic acid fragment is capable significantly reducing the amount of DCL-protein of SEQ ID NO:3 or SEQ ID NO:4 in a cancer cells of neuroectodermal origin when said fragment is introduced into said cell.
6 . A composition comprising one or more nucleic acid fragments according to claim 5 and a physiologically acceptable carrier.
7 . The composition according to claim 6 , further comprising one or more targeting compounds that target cancer cells of neuroectodermal origin in vivo or in vitro.
8 . The composition according to claim 7 , wherein the targeting compound is an immunoliposome or a monoclonal antibody specific for said cancer cells.
9 . The composition according to claim 6 , wherein said composition is suitable for treatment of a cancer of neuroectodermal origin.
10 . A doublecortin-like protein, the sequence of which is SEQ ID NO:3 or SEQ ID NO:4.
11 . A method for diagnosing cancers of neuroectodermal origin, comprising the steps of
(a) analyzing a biological sample from a subject for the presence or absence of:
(i) DNA or RNA, the sequence of which is SEQ ID NO:2 and/or
(ii) DCL protein, the sequence of which is SEQ ID NO:4; and optionally
(b) quantifying the amount of said DNA, RNA or protein in the sample,
wherein the presence of said DNA, RNA or protein is indicative of the presence of the cancer in the subject.
12 . A diagnostic kit for diagnosing cancer of neuroectodermal origin in a biological sample from a mammalian subject, said kit comprising one or more primers, probes and/or antibodies that hybridize to, or bind to, and thereby detect the presence of a nucleic acid with a sequence SEQ ID NO:2 or a polypeptide with a sequence SEQ ID NO:4, and instructions for use of said kit.
13 . The method according to claim 4 wherein the nucleic acid fragment is an antisense RNA oligonucleotide selected from the group consisting of SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15 and SEQ ID NO:16.
14 . The method according to claim 4 wherein the nucleic acid fragment is an antisense DNA oligonucleotide selected from the group consisting of SEQ ID NO: 17, SEQ ID NO:18, SEQ ID NO:19 and SEQ ID NO:20.
15 . The method according to claim 4 wherein the nucleic acid fragment is a double stranded small interfering RNA oligonucleotide selected from the group consisting of double stranded sequences
SEQ ID NO:5/SEQ ID NO:6,
SEQ ID NO:7/SEQ ID NO:8,
SEQ ID NO:9/SEQ ID NO:10 and
SEQ ID NO:11/SEQ ID NO:12.
16 . The nucleic acid fragment according to claim 5 which is an antisense RNA oligonucleotide selected from the group consisting of SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15 and SEQ ID NO:16.
17 . The nucleic acid fragment according to claim 5 which is an antisense DNA oligonucleotide selected from the group consisting of SEQ ID NO: 17, SEQ ID NO:18, SEQ ID NO:19 and SEQ ID NO:20
18 . The nucleic acid fragment according to claim 5 which is a double stranded small interfering RNA oligonucleotide selected from the group consisting of double stranded sequences
SEQ ID NO:5/SEQ ID NO:6,
SEQ ID NO:7/SEQ ID NO:8,
SEQ ID NO:9/SEQ ID NO:10 and
SEQ ID NO:11/SEQ ID NO:12.
19 . A method according to claim 11 wherein said quantification step (b) is performed, and wherein the amount of said protein, DNA or RNA is correlated to the number cancer cells of neuroectodermal origin present in the subject and to the severity of the cancer.
20 . A method of inhibiting the growth of cancer cells of neuroectodermal origin, comprising providing to the cells an effective amount of a nucleic acid fragment of SEQ ID NO:1 or SEQ ID NO:2, or a fragment of a variant of SEQ ID NO:1 or SEQ ID NO:2, which nucleic acid fragment significantly reduces the amount of a DCL-protein, the sequence of which is SEQ ID NO:3 or SEQ ID NO:4, in said cells, thereby inhibiting their growth.