Mir-211 expression and related pathways in human melanoma

a human melanoma and gene expression technology, applied in the field of human melanoma diagnosis and treatment, can solve the problems of complex and poorly understood exact molecular mechanisms that lead to melanoma, significant hurdle to its use, and the extent of melanin pigmentation is an important risk factor

Inactive Publication Date: 2014-05-15
SANFORD BURNHAM MEDICAL RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

A major cause for melanoma initiation is extensive or intermittent exposure to the sun's radiation over a period of time, and the extent of melanin pigmentation is an important risk factor.
The exact molecular mechanisms that lead to melanoma are complex and poorly understood, and may involve both mutagenic DNA lesions and epigenetic misregulation.
The loss of 1NK4a therefore may lead to interference of two separate pathways of cell cycle control: CDK signaling and suppression of p53 activity by Mdm2-induced acceleration of p53 degradation.
Several small-molecule inhibitors of BRAF are known, such as vemurafenib, but a significant hurdle to its use exists due to the emergence of vemurafenib-resistant cells.

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  • Mir-211 expression and related pathways in human melanoma
  • Mir-211 expression and related pathways in human melanoma
  • Mir-211 expression and related pathways in human melanoma

Examples

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example 1

miR-211 is Expressed at a Low Level in Non-Pigmented Melanoma Cell Lines

[0104]The human epidermal melanocyte cell line HEM-1 (ScienCell™, Catalog #2200) and primary epidermal melanoyctes—neonatal (ATCC-PCS-200-012) were grown in Me1M media containing MeIGS growth supplements, 0.5% FBS, and pen / strep solution. The melanoma cell lines examined included: A375 (stage 4, ATCC® Number: CRL-1619), G361 (stage 4, ATCC), LOX-IMV1 (stage 4, ATCC), HT-144 (stage 4, ATCC® Number: HTB-63), RPMI-7951 (stage 4, ATCC® Number: HTB-66), SK-MEL2 (stage 4, ATCC), SK-MEL28 (stage 3, ATCC), WM793B (stage 1, ATCC® Number: CRL-2806), and WM1552C (stage 3, ATCC® Number: CRL-2808). All melanoma cell lines were grown in Complete Tu Media containing a 4:1 mixture of MCDB-153 medium with 1.5 g / L sodium bicarbonate and Leibovitz's L-15 medium with 2 mM L-glutamine, 2% FBS, and 1.68 mM CaCl2. Information regarding all clinical samples, derived from frozen samples, is described in Table 1.

TABLE 1Clinical Sample #T...

example 2

miR-211 Levels in Clinical Melanoma Samples

[0108]miR-211 transcript levels were assayed by qRT-PCR in 30 clinical melanoma samples (six primary, six regional, 12 nodal and six distal metastatic, respectively; described in Table S1). miR-211 expression levels were reduced in 21 of these clinical samples compared to that observed in melanocytes (FIG. 3, Table 3). In the remaining nine melanomas, six (one primary, one regional, two distant, and two nodal metastatic melanomas) showed statistically significant increases in miR-211 expression, whereas expression was not significantly different in the remaining samples. These samples were obtained from different patients; therefore, the observed differences may reflect different processes in melanoma development and progression, individual genetic differences, different proportions of non-melanoma (including non-pigmented) cells in the tumor samples, or a combination of these factors. miR-211 levels were low in the majority (21 / 30) of the ...

example 3

Stable Ectopic Expression of miR-211 in Melanoma Cell Lines Depletes Select Target Transcripts

[0110]For the initial transformation of miRNA array data, the GenePixPro 6.0 global normalization method was employed in which images and results are normalized together. Statistical significance tests were Welsh t-test, nonparametric ANOVA, (e.g., to select genes that have significantly less within sample variance compared to between sample variance), and correlation analysis with Pearson's product moment r and Spearman's r. Analysis was controlled for false discovery rate using q-values, with a priori cut off point of 10 percent

[0111]For mRNA expression array data, commencing with GeneChip® Human Exon 1.0 ST Array (Affymetrix, Inc.) four probes per exon and roughly 40 probes per gene, 7 total arrays were analysed (three arrays for melanocyte RNA, and four arrays for melanoma RNA). Cell files were loaded into Partek® Genomics Suite™ (Partek, Inc. St. Louis, Mo., USA) under the following al...

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Abstract

Provided herein are methods for the diagnosis of resistance to chemotherapeutic agents by assessing the regulatory pathways of PGC1α. Methods for treating drug-resistant melanoma are also provided.

Description

CROSS REFERENCE TO RELATED APPLICATIONS[0001]This application is a continuation-in-part of U.S. patent application Ser. No. 13 / 271,030, filed Oct. 11, 2011 and published as US 2012 / 0108457, which claims priority to U.S. Provisional Patent Application No. 61 / 391,948, filed Oct. 11, 2010 and 61 / 442,108, filed Feb. 11, 2011, each of which is incorporated by reference in its entirety.FIELD OF THE INVENTION[0002]The present invention relates to methods of diagnosing and treating human melanoma characterized by resistance to at least one chemotherapeutic agent.BACKGROUND OF THE INVENTION[0003]The following discussion of the background of the invention is merely provided to aid the reader in understanding the invention and is not admitted to describe or constitute prior art to the present invention.[0004]Melanoma, a cancer of the pigment-producing cells in the skin epidermis, can be highly metastatic, and malignant melanomas are relatively resistant to standard chemotherapy. A major cause ...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12N15/113A61K38/17C12Q1/68
CPCC12N15/113C12Q1/6881A61K38/1709C12Q1/6886C12N15/1138C12N2310/14C12N2310/141C12N2310/531C12N2320/10C12Q2600/106C12Q2600/158C12Q2600/178G01N33/5743
Inventor PERERA, RANJAN
Owner SANFORD BURNHAM MEDICAL RES INST
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