MicroRNA Expression Abnormalities in Pancreatic Endocrine and Acinar Tumors

Abstract

The present invention provides novel methods and compositions for the diagnosis, prognosis and treatment of pancreatic cancer. The invention also provides methods of identifying anti-pancreatic cancer agent.

What is claimed is: 
     
         1 . A method of determining whether a subject has, or is at risk for developing, pancreatic cancer, wherein the method comprises:
 extracting from the subject a test sample comprising at a miR-103 gene product;   measuring the level of the gene product in the test sample;   determining whether the subject has, or is at risk for developing, pancreatic cancer by comparing the level of the gene product in the test sample to the level of a corresponding miR gene product in a control sample; and,   correlating an increase in the level of the miR-103 gene product in the test sample, relative to the level of the corresponding miR gene product in the control sample, as being indicative of a diagnosis of the subject either having, or being at risk for developing, pancreatic cancer.   
     
     
         2 . The method according to  claim 1 , wherein the test sample further comprises at least one miR-107 gene product, and wherein the method further comprises:
 measuring the level of the miR-107 gene product in the test sample;   determining whether the subject has, or is at risk for developing, pancreatic cancer by comparing the level of the miR-107 gene product in the test sample to the level of a corresponding miR gene product in a control sample; and,   correlating an increase in the level of the miR-107 gene product in the test sample, relative to the level of the corresponding miR gene product in the control sample, as being indicative of a diagnosis of the subject either having, or being at risk for developing, pancreatic cancer.   
     
     
         3 . The method according to  claim 1 , wherein the test sample further comprises at least one additional gene product selected from miR-23a, miR-26b, miR-192 and miR-342, and wherein the method further comprises:
 measuring the level of the at least one additional miR gene product in the test sample;   determining whether the subject has, or is at risk for developing, pancreatic cancer by comparing the level of the at least one additional miR gene product in the test sample to the level of a corresponding miR gene product in a control sample; and,   correlating an increase in the level of the at least one additional miR gene product in the test sample, relative to the level of the corresponding miR gene product in the control sample, as being indicative of a diagnosis of the subject either having, or being at risk for developing, pancreatic cancer.   
     
     
         4 . The method according to  claim 1 , wherein the test sample further comprises at least miR-107 gene product and at least one additional gene product selected from miR-23a, miR-26b, miR-192 and miR-342, and wherein the method further comprises:
 measuring the level of the miR-107 gene product and the level of the at least one additional miR gene product in the test sample;   determining whether the subject has, or is at risk for developing, pancreatic cancer by comparing the level of the miR-107 gene product and the level of the at least one additional gene product in the test sample to the level of a corresponding miR gene product in a control sample; and   correlating an increase in the level of the miR-107 gene product and an increase in the level of the at least one additional miR gene product in the test sample, relative to the level of the corresponding miR gene product in the control sample, as being indicative of a diagnosis of the subject either having, or being at risk for developing, pancreatic cancer.   
     
     
         5 . The method according to  claim 1 , wherein the measuring the level of the miR-103 gene product in the test sample is carried out by analyzing the test sample with a RT-PCR, a microarray and/or a Northern blot electrophoretic device. 
     
     
         6 . The method according to  claim 2 , wherein the measuring the level of the miR-103 gene product and the level of the miR-103 gene product in the test sample is carried out by analyzing the test sample with a RT-PCR, a microarray and/or a Northern blot electrophoretic device. 
     
     
         7 . The method according to  claim 3 , wherein the measuring the level of the miR-103 gene product and the level of the at least one additional gene product in the test sample is carried out by analyzing the test sample with a RT-PCR, a microarray and/or a Northern blot electrophoretic device. 
     
     
         8 . The method according to  claim 4 , wherein the measuring the level of the miR-103 gene product, the level of the miR-107 gene product, and the level of the at least one additional gene product in the test sample is carried out by analyzing the test sample with a RT-PCR, a microarray and/or a Northern blot electrophoretic device. 
     
     
         9 . The method according to  claim 1 , further comprising: confirming diagnosis by searching, or periodically monitoring, for symptoms which are associated with pancreatic cancer and exhibited by the subject when there is an increase in the level of the miR-103 gene product in the test sample, relative to the level of the corresponding miR gene product in the control sample. 
     
     
         10 . The method according to  claim 2 , further comprising: confirming diagnosis by searching, or periodically monitoring, for symptoms which are associated with pancreatic cancer and exhibited by the subject when there is an increase in the level of the miR-103 gene product in the test sample, relative to the level of the corresponding miR gene product in the control sample. 
     
     
         11 . The method according to  claim 3 , further comprising: confirming diagnosis by searching, or periodically monitoring, for symptoms which are associated with pancreatic cancer and exhibited by the subject when there is an increase in the level of the miR-103 gene product and an increase in the level of the miR-107 gene product in the test sample, relative to the level of the corresponding miR gene product in the control sample. 
     
     
         12 . The method according to  claim 4 , further comprising: confirming diagnosis by searching, or periodically monitoring, for symptoms which are associated with pancreatic cancer and exhibited by the subject when there is an increase in the level of the miR-103 gene product, an increase in the level of the miR-107 gene product, and an increase in the level of the at least one additional miR gene product in the test sample, relative to the level of the corresponding miR gene product in the control sample. 
     
     
         13 . The method according to  claim 1 , wherein the pancreatic cancer is a pancreatic endocrine tumor (PET). 
     
     
         14 . The method according to  claim 13 , wherein the pancreatic cancer is at least one pancreatic endocrine tumor (PET) selected from the group consisting of gastrinoma, insulinoma, somatostatinoma, VIPoma, and glucagonoma. 
     
     
         15 . The method according to  claim 14 , further comprising: confirming diagnosis by searching, or periodically monitoring, for symptoms of hormone hypersecretion associated with PET and exhibited by the subject when there is an increase in the level of the miR-103 gene product in the test sample, relative to the level of the corresponding miR gene product in the control sample. 
     
     
         16 . The method according to  claim 15 , wherein the hormone is at least one hypersecreted hormone selected from the group consisting of gastrin, insulin, somatostatin, vasoactive intestinal polypeptide (VIP), and glucagon. 
     
     
         17 . The method according to  claim 2 , wherein the pancreatic cancer is a pancreatic endocrine tumor (PET). 
     
     
         18 . The method according to  claim 17 , wherein the pancreatic cancer is at least one pancreatic endocrine tumor (PET) selected from the group consisting of gastrinoma, insulinoma, somatostatinoma, VIPoma, and glucagonoma. 
     
     
         19 . The method according to  claim 18 , further comprising: confirming diagnosis by searching, or periodically monitoring, for symptoms of hormone hypersecretion associated with PET and exhibited by the subject when there is an increase in the level of the miR-103 gene product and an increase in the level of the miR-107 gene product in the test sample, relative to the level of the corresponding miR gene product in the control sample. 
     
     
         20 . The method according to  claim 19 , wherein the hormone is at least one hypersecreted hormone selected from the group consisting of gastrin, insulin, somatostatin, vasoactive intestinal polypeptide (VIP), and glucagon. 
     
     
         21 . The method according to  claim 3 , wherein the pancreatic cancer is a pancreatic endocrine tumor (PET). 
     
     
         22 . The method according to  claim 21 , wherein the pancreatic cancer is at least one pancreatic endocrine tumor (PET) selected from the group consisting of gastrinoma, insulinoma, somatostatinoma, VIPoma, and glucagonoma. 
     
     
         23 . The method according to  claim 22 , further comprising: confirming diagnosis by searching, or periodically monitoring, for symptoms of hormone hypersecretion associated with PET and exhibited by the subject when there is an increase in the level of the miR-103 gene product and an increase in the level of the at least one additional miR gene product in the test sample, relative to the level of the corresponding miR gene product in the control sample. 
     
     
         24 . The method according to  claim 23 , wherein the hormone is at least one hypersecreted hormone selected from the group consisting of gastrin, insulin, somatostatin, vasoactive intestinal polypeptide (VIP), and glucagon. 
     
     
         25 . The method according to  claim 4 , wherein the pancreatic cancer is a pancreatic endocrine tumor (PET). 
     
     
         26 . The method according to  claim 25 , wherein the pancreatic cancer is at least one pancreatic endocrine tumor (PET) selected from the group consisting of gastrinoma, insulinoma, somatostatinoma, VIPoma, and glucagonoma. 
     
     
         27 . The method according to  claim 26 , further comprising: confirming diagnosis by searching, or periodically monitoring, for symptoms of hormone hypersecretion associated with PET and exhibited by the subject when there is an increase in the level of the miR-103 gene product, an increase in the level of the miR-107 gene product, and an increase in the level of the at least one additional miR gene product in the test sample, relative to the level of the corresponding miR gene product in the control sample. 
     
     
         28 . The method according to  claim 27 , wherein the hormone is at least one hypersecreted hormone selected from the group consisting of gastrin, insulin, somatostatin, vasoactive intestinal polypeptide (VIP), and glucagon. 
     
     
         29 . The method according to  claim 1 , further comprising: communicating to the subject the diagnosis of having, or being at risk for developing, pancreatic cancer when there is an increase in the level of the miR-103 gene product in the test sample, relative to the level of the corresponding miR gene product in the control sample. 
     
     
         30 . The method according to  claim 2 , further comprising: communicating to the subject the diagnosis of having, or being at risk for developing, pancreatic cancer when there is an increase in the level of the miR-103 gene product and an increase in the level of the miR-107 gene product in the test sample, relative to the level of the corresponding miR gene product in the control sample. 
     
     
         31 . The method according to  claim 3 , further comprising: communicating to the subject the diagnosis of having, or being at risk for developing, pancreatic cancer when there is an increase in the level of the miR-103 gene product and an increase in the level of the at least one additional miR gene product in the test sample, relative to the level of the corresponding miR gene product in the control sample. 
     
     
         32 . The method according to  claim 4 , further comprising: communicating to the subject the diagnosis of having, or being at risk for developing, pancreatic cancer when there is an increase in the level of the at least miR-103 gene product, an increase in the level of the miR-107 gene product, and an increase in the level of the at least one additional miR gene product in the test sample, relative to the level of the corresponding miR gene product in the control sample. 
     
     
         33 . The method according to  claim 29 , further comprising: communicating to the subject the diagnosis of having, or being at risk for developing, PET when there is an increase in the level of the miR-103 gene product in the test sample, relative to the level of the corresponding miR gene product in the control sample, and symptoms of hormone hypersecretion associated PET are exhibited by the subject. 
     
     
         34 . The method according to  claim 30 , further comprising: communicating to the subject the diagnosis of having, or being at risk for developing, PET when there is an increase in the level of the miR-103 gene product and an increase in the level of the miR-107 gene product in the test sample, relative to the level of the corresponding miR gene product in the control sample, and symptoms of hormone hypersecretion associated PET are exhibited by the subject. 
     
     
         35 . The method according to  claim 31 , further comprising: communicating to the subject the diagnosis of having, or being at risk for developing, PET when there is an increase in the level of the miR-103 gene product and an increase in the level of the at least one additional miR gene product in the test sample, relative to the level of the corresponding miR gene product in the control sample, and symptoms of hormone hypersecretion associated PET are exhibited by the subject. 
     
     
         36 . The method according to  claim 32 , further comprising: communicating to the subject the diagnosis of having, or being at risk for developing, PET when there is an increase in the level of the at least miR-103 gene product, an increase in the level of the miR-107 gene product, and an increase in the level of the at least one additional miR gene product in the test sample, relative to the level of the corresponding miR gene product in the control sample, and symptoms of hormone hypersecretion associated PET are exhibited by the subject. 
     
     
         37 . A method of determining whether a subject has, or is at risk for developing, pancreatic cancer, wherein the method comprises:
 isolating RNA from a test sample extracted from the subject;   reverse transcribing the RNA isolated from the test sample to provide at least one target oligodeoxynucleotide;   hybridizing the at least one target oligodeoxynucleotide to a microarray comprising at least one miR-103 miRNA-specific probe oligonucleotide to provide a hybridization profile for the test sample;   determining whether the subject has, or is at risk for developing, pancreatic cancer by comparing the signal of the miR-103 miRNA in the hybridization profile for the test sample to the signal of a corresponding miRNA in the hybridization profile for a control sample; and,   correlating a up-regulated signal of the miR-103 miRNA in the test sample, relative to the signal of the corresponding miRNA in the control sample, as being indicative of a diagnosis of the subject either having, or being at risk for developing, pancreatic cancer.   
     
     
         38 . The method according to  claim 37 , wherein the method further comprises:
 hybridizing the target oligodeoxynucleotide to the microarray which further comprises at least one miR-107 miRNA-specific probe oligonucleotide to provide a hybridization profile for the test sample;   determining whether the subject has, or is at risk for developing, pancreatic cancer by comparing the signal of the miR-107 miRNA in the hybridization profile for the test sample to the signal of a corresponding miRNA in the hybridization profile for a control sample; and,   correlating an upregulated signal of the miR-107 miRNA in the test sample, relative to the signal of the corresponding miRNA in the control sample, as being indicative of a diagnosis of the subject either having, or being at risk for developing, pancreatic cancer.   
     
     
         39 . The method according to  claim 37 , wherein the method further comprises:
 hybridizing the target oligodeoxynucleotide to the microarray which further comprises at least one additional miR miRNA-specific probe oligonucleotide to provide a hybridization profile for the test sample;   determining whether the subject has, or is at risk for developing, pancreatic cancer by comparing the signal of the at least one additional miR miRNA in the hybridization profile for the test sample to the signal of a corresponding miRNA in the hybridization profile for a control sample; and,   correlating an upregulated signal of the at least one additional miR miRNA in the test sample, relative to the signal of the corresponding miRNA in the control sample, as being indicative of a diagnosis of the subject either having, or being at risk for developing, pancreatic cancer.   
     
     
         40 . The method according to  claim 37 , wherein the method further comprises:
 hybridizing the target oligodeoxynucleotide to the microarray which further comprises at least one miR-107 miRNA-specific probe oligonucleotide and at least one additional miR miRNA-specific probe oligonucleotide to provide a hybridization profile for the test sample;   determining whether the subject has, or is at risk for developing, pancreatic cancer by comparing the signal of the miR-107 miRNA and the signal of the at least one additional miR miRNA in the hybridization profile for the test sample to the signal of a corresponding miRNA in the hybridization profile for a control sample; and,   correlating an upregulated signal of the miR-107 miRNA and an upregulated signal of the at least one additional miRmiRNA in the test sample, relative to the signal of the corresponding miRNA in the control sample, as being indicative of a diagnosis of the subject either having, or being at risk for developing, pancreatic cancer.   
     
     
         41 . The method according to  claim 37 , further comprising: confirming diagnosis by searching, or periodically monitoring, for symptoms which are associated with pancreatic cancer and exhibited by the subject when there is an upregulated signal of the miR-103 miRNA in the test sample, relative to the signal of the corresponding miRNA in the control sample. 
     
     
         42 . The method according to  claim 38 , further comprising: confirming diagnosis by searching, or periodically monitoring, for symptoms which are associated with pancreatic cancer and exhibited by the subject when there is an upregulated signal of the miR-103 miRNA and an upregulated signal of the miR-107 miRNA in the test sample, relative to the signal of the corresponding miRNA in the control sample. 
     
     
         43 . The method according to  claim 39 , further comprising: confirming diagnosis by searching, or periodically monitoring, for symptoms which are associated with pancreatic cancer and exhibited by the subject when there is an upregulated signal of the miR-103 miRNA and an upregulated signal of the at least one additional miR miRNA in the test sample, relative to the signal of the corresponding miRNA in the control sample. 
     
     
         44 . The method according to  claim 40 , further comprising: confirming diagnosis by searching, or periodically monitoring, for symptoms which are associated with pancreatic cancer and exhibited by the subject when there is an upregulated signal of the miR-103 miRNA, an upregulated signal of the miR-107 miRNA, and an upregulated signal of the at least one additional miR miRNA in the test sample, relative to the signal of the corresponding miRNA in the control sample. 
     
     
         45 . The method according to  claim 37 , wherein the pancreatic cancer is a pancreatic endocrine tumor (PET). 
     
     
         46 . The method according to  claim 37 , wherein the pancreatic cancer is at least one pancreatic endocrine tumor (PET) selected from the group consisting of gastrinoma, insulinoma, somatostatinoma, VIPoma, and glucagonoma. 
     
     
         47 . The method according to  claim 46 , further comprising: confirming diagnosis by searching, or periodically monitoring, for symptoms of hormone hypersecretion associated with PET and exhibited by the subject when there is an upregulated signal of the miR-103 miRNA in the test sample, relative to the signal of the corresponding miRNA in the control sample. 
     
     
         48 . The method according to  claim 47 , wherein the hormone is at least one hypersecreted hormone selected from the group consisting of gastrin, insulin, somatostatin, vasoactive intestinal polypeptide (VIP), and glucagon. 
     
     
         49 . The method according to  claim 38 , wherein the pancreatic cancer is a pancreatic endocrine tumor (PET). 
     
     
         50 . The method according to  claim 49 , wherein the pancreatic cancer is at least one pancreatic endocrine tumor (PET) selected from the group consisting of gastrinoma, insulinoma, somatostatinoma, VIPoma, and glucagonoma. 
     
     
         51 . The method according to  claim 50 , further comprising: confirming diagnosis by searching, or periodically monitoring, for symptoms of hormone hypersecretion associated with PET and exhibited by the subject when there is an upregulated signal of the miR-103 miRNA and an upregulated signal of the miR-107 miRNA in the test sample, relative to the signal of the corresponding miRNA in the control sample. 
     
     
         52 . The method according to  claim 51 , wherein the hormone is at least one hypersecreted hormone selected from the group consisting of gastrin, insulin, somatostatin, vasoactive intestinal polypeptide (VIP), and glucagon. 
     
     
         53 . The method according to  claim 39 , wherein the pancreatic cancer is a pancreatic endocrine tumor (PET). 
     
     
         54 . The method according to  claim 53 , wherein the pancreatic cancer is at least one pancreatic endocrine tumor (PET) selected from the group consisting of gastrinoma, insulinoma, somatostatinoma, VIPoma, and glucagonoma. 
     
     
         55 . The method according to  claim 54 , further comprising: confirming diagnosis by searching, or periodically monitoring, for symptoms of hormone hypersecretion associated with PET and exhibited by the subject when there is an upregulated signal of the miR-103 miRNA and an upregulated signal of the at least one additional miR miRNA in the test sample, relative to the signal of the corresponding miRNA in the control sample. 
     
     
         56 . The method according to  claim 55 , wherein the hormone is at least one hypersecreted hormone selected from the group consisting of gastrin, insulin, somatostatin, vasoactive intestinal polypeptide (VIP), and glucagon. 
     
     
         57 . The method according to  claim 40 , wherein the pancreatic cancer is a pancreatic endocrine tumor (PET). 
     
     
         58 . The method according to  claim 57 , wherein the pancreatic cancer is at least one pancreatic endocrine tumor (PET) selected from the group consisting of gastrinoma, insulinoma, somatostatinoma, VIPoma, and glucagonoma. 
     
     
         59 . The method according to  claim 58 , further comprising: confirming diagnosis by searching, or periodically monitoring, for symptoms of hormone hypersecretion associated with PET and exhibited by the subject when there is an upregulated signal of the miR-103 miRNA, an upregulated signal of the miR-107 miRNA, and an upregulated signal of the at least one additional miR miRNA in the test sample, relative to the signal of the corresponding miRNA in the control sample. 
     
     
         60 . The method according to  claim 59 , wherein the hormone is at least one hypersecreted hormone selected from the group consisting of gastrin, insulin, somatostatin, vasoactive intestinal polypeptide (VIP), and glucagon. 
     
     
         61 . The method according to  claim 37 , further comprising: communicating to the subject the diagnosis of having, or being at risk for developing, pancreatic cancer when there is an upregulated signal of the miR-103 miRNA in the test sample, relative to the signal of the corresponding miRNA in the control sample. 
     
     
         62 . The method according to  claim 38 , further comprising: communicating to the subject the diagnosis of having, or being at risk for developing, pancreatic cancer when there is an upregulated signal of the miR-103 miRNA and an upregulated signal of the miR-107 miRNA in the test sample, relative to the signal of the corresponding miRNA in the control sample. 
     
     
         63 . The method according to  claim 39 , further comprising: communicating to the subject the diagnosis of having, or being at risk for developing, pancreatic cancer when there is an upregulated signal of the miR-103 miRNA and an upregulated signal of the at least one additional miR miRNA in the test sample, relative to the signal of the corresponding miRNA in the control sample. 
     
     
         64 . The method according to  claim 40 , further comprising: communicating to the subject the diagnosis of having, or being at risk for developing, pancreatic cancer when there is an upregulated signal of the miR-103 miRNA, an upregulated signal of the miR-107 miRNA and an upregulated signal of the at least one additional miR miRNA in the test sample, relative to the signal of the corresponding miRNA in the control sample. 
     
     
         65 . The method of  claim 3 , wherein the type of pancreatic cancer is a pancreatic endocrine tumor (PET), and the at least one additional miR gene product is further selected from the group consisting of: miR-125a, miR-99a, miR-99b, miR-125b-1, miR-342, miR-130a, miR-100, miR-132, miR-129-2, miR-125b-2 and a combination thereof. 
     
     
         66 . The method of  claim 1 , wherein the miR gene product comprises miR-103 and wherein the increased presence of miR-103 discriminates among normal pancreas, acinar cell carcinomas and pancreatic endocrine tumors. 
     
     
         67 . A probe for hybridization comprising a miRNA selected from the group consisting of a sequence that is complementary to a miRNA selected from miR-103, miR-107, miR-125a, miR-99a, miR-99b, miR-125b-1, miR-342, miR-130a, miR-100, miR-132, miR-129-2, miR-125b-2; a fragment thereof and a sequence having at least about 80% identity thereto. 
     
     
         68 . The probe of  claim 67 , wherein the cancer is pancreatic endocrine cancer.