Compositions and methods for diagnosis and prognosis of cancer and progression, and for screening anti-cancer agents
MicroRNA (miRNA) profiling of cells showed unique miRNA signatures for each of three Akt isoforms. Among differentially regulated miRNA species, the miR-200 family was downregulated in Akt2-expressing cells. Akt1 knockdown inhibited expression of miR-200 and promoted TGFβ-induced epithelial-mesenchymal-transition (EMT) and a stem cell like phenotype. Carcinomas developing in MMTV-cErb2/Akt1 −/− mice exhibited increased invasiveness because of EMT induced by miR-200 downregulation. EMT was found to be controlled by miRNA species that are regulated by the balance between Akt1 and Akt2, rather than overall Akt levels.
1. A method for prognosing or diagnosing a potential cancer and progression of an existing cancer in a patient by analyzing Akt isoform imbalance, wherein the imbalance comprises a decreased ratio of Akt1 protein to Akt2 protein compared to that ratio in a normal subject, the method comprising:
measuring an expression profile of a plurality of microRNA (miRNA) species regulated by Akt isoform expression in a sample from the patient, by real-time PCR, or by using an array of miRNA nucleotide sequences of a plurality of miRNA species each sequence at a known location on a substrate; and
comparing the profile from the patient with that of the normal subject not having the cancer and obtaining an miRNA tumor expression profile;
wherein the decreased ratio of the Akt isoform expression profile of the sample from the patient compared to the normal subject is a prognosis or diagnosis of the potential cancer or an indication of the progression of the existing cancer in the patient.
2. The method according to claim 1 , wherein the miRNA is miR-200a.
3. The method according to claim 1 , wherein the sample comprises a cell from a biopsy of the patient comprising at least one selected from the groupage of: blood, cerebrospinal fluid, mammary glands, prostate gland, lung, bladder, cervix, and colorectal, the method further comprising culturing cells ex vivo and screening compounds in a library contacted with aliquots of resulting cultured cells and control normal subject cells, to obtain a potential inhibitor that restores Akt expression balance to an amount observed for the normal subject and thereby inhibits the cancer.
4. The method according to claim 1 , further comprising measuring decreased expression compared to the normal subject of at least one transcription factor miRNA, and prognosing the cancer or the cancer progression from the decreased transcription factor expression of the transcription factor.
5. The method according to claim 4 , wherein the transcriptional factor is selected from at least one of: IGF1 and TGFβ.
6. The method according to claim 1 , further comprising measuring Akt amounts and observing no difference between amounts in patient and in normal subject Akt amounts, and correlating with at least one selected from: the cancer, the progression of the cancer, metastasis, invasiveness, enlargement of tumor size, regression of tumor size, disappearance of tumor, no evidence of disease, and remission.
7. The method according to claim 1 , further comprising determining cell motility of the sample, and prognosing increased motility with at least one of invasiveness and metastasis in the patient.
8. The method according to claim 1 , further comprising measuring upregulation of expression of at least one miRNA in hypoxic cells or in a hypoxic tumor.
9. The method according to claim 1 , further comprising measuring decreased expression of miR-200a and prognosing the cancer or the cancer progression in the patient.
10. The claim according to claim 9 , further comprising measuring decreased expression of at least one miRNA regulated by the imbalance of Akt isoforms having the decreased ratio of the Akt1 protein to the Akt2 protein compared to the normal subject, and prognosing the cancer or the cancer progression in the patient.
11. The method according to claim 10 , further comprising measuring the decreased expression of the miRNA species regulated by the imbalance of expression of at least one of the Akt1 protein and the Akt2 protein, and prognosing the cancer or the cancer progression in the patient.
12. The method according to claim 11 , further comprising measuring the miRNA species regulated by the reduced expression of the Akt1 protein, and prognosing the cancer or the cancer progression in the patient.
13. The method according to claim 1 , further comprising measuring increased expression of the miRNA species regulated by the imbalance of at least one of the Akt1 protein, the Akt2 protein and an Akt3 protein, and prognosing the cancer or the cancer progression in the patient.