Methods and kits for detecting subjects at risk of having cancer

#This invention relates to methods and agents for high risk screening, early diagnosis and surveillance of cancer, in particular pancreatic cancer.

#Cancer, also known as a malignant neoplasm, refers to disorders involving unregulated cell growth. In cancer, cells divide and grow uncontrollably, forming malignant tumors, and invade nearby parts of the body. The cancer may spread to distant parts of the body through the lymphatic system or bloodstream. Despite significant advances in therapeutics and diagnostics, cancer remains a major cause of morbidity and mortality in the U.S. For some cancers, despite these advances, the incidence is on the rise.

#For example, pancreatic cancer represents an aggressive tumor with less than a 5% survival rate (Jemal et al., Cancer statistics CA Cancer J. Clin. 2009; 59:225-249). Recent findings from Japan indicate that, with small tumors and no local infiltration, the survival rate can improve to over 20% (Tanaka et al. Pancreas 2004; 28(3):268-72). Survival is better upon early diagnosis. Unfortunately, the majority of pancreatic cancer patients (85%) are diagnosed at late stage as diagnosis of pancreatic cancer at an early stage has met with several challenges including lack of biomarkers suitable for early detection and associated non-cancerous pancreatic diseases, which complicate early detection. Many molecules from different classes have been interrogated as potential early detection markers, but yield no success.

#FIG. 1 is a diagram showing results of unsupervised cluster analysis revealed a significant (FDR<10%) and differentially regulated (>2-fold) miRNA signature comprising of 14 members that segregated pancreatic cancer patients away from not only healthy individuals but also high-risk subjects. These 14 significant microRNAs were identified from microarray experiments that probed the entire Sanger miRBASE version 16. MicroRNA probe expression values (Log 2 transformed & normalized microarray probe intensities) of selected microRNA in Cancer, High Risk and Control samples were median centered. Each column represents a single sample, and each row represents a single microRNA probe. Green squares represent lower than median levels of microRNA expression; black squares represent median levels of microRNA expression; red squares represent higher than median levels of microRNA expression. Legend units: 1.0=differs from median probe intensity by one log 2 unit (2-fold)

#This invention is based, at least in part, on an unexpected discovery of a number of microRNA gene products whose expression levels are altered in biological samples obtained from subjects with cancer, such as pancreatic cancer, relative to control samples. These genes can be used as biomarkers for determining whether a subject has, or is at risk of having, a cellular proliferative disorder or for determining a prognosis or surveillance of patient having such a disorder.

#Accordingly, the present invention encompasses methods of diagnosing whether a subject has, or is at risk for, a cellular proliferative disorder, such as cancer or neoplastic diseases. The term “neoplastic diseases” refers to cancers of any kind and origin and precursor stages thereof. The term “neoplastic disease” includes the subject matter identified by the terms “neoplasia,” “neoplasm,” “cancer,” “pre-cancer,” or “tumor.” A neoplastic disease is generally manifest by abnormal cell division resulting in an abnormal level of a particular cell population. The abnormal cell division underlying a neoplastic disease is typically inherent in the cells and not a normal physiological response to infection or inflammation. In some embodiments, neoplastic diseases for diagnosis using methods provided herein include carcinoma. By “carcinoma,” it is meant a benign or malignant epithelial tumor and includes, but is not limited to, hepatocellular carcinoma, breast carcinoma, prostate carcinoma, non-small cell lung carcinoma, colon carcinoma, CNS carcinoma, melanoma, ovarian carcinoma, or renal carcinoma. An exemplary neoplastic disease is pancreatic cancer, including adenocarcinoma and neuroendocrine tumor.

#The invention also provides for methods of screening subjects who are thought to be at risk for developing the above-mentioned cancer, e.g., pancreatic cancer. Also provided are methods of determining the efficacy of therapeutic regimens for inhibiting the cancer, and methods of identifying an anti-cancer agent. The invention also encompasses various kits suitable for carrying out the above mentioned methods.