Use of FGFR mutant gene panels in identifying cancer patients that will be responsive to treatment with an FGFR inhibitor

A technology for FGFR3G370C, FGFR3, applied in the field of using FGFR mutant genome to identify cancer patients who will respond to treatment with FGFR inhibitors, can solve problems such as patients without FGFR changes

Active Publication Date: 2017-08-01
JANSSEN PHARMA NV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There are currently no approved therapies that ar

Method used

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  • Use of FGFR mutant gene panels in identifying cancer patients that will be responsive to treatment with an FGFR inhibitor
  • Use of FGFR mutant gene panels in identifying cancer patients that will be responsive to treatment with an FGFR inhibitor
  • Use of FGFR mutant gene panels in identifying cancer patients that will be responsive to treatment with an FGFR inhibitor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0199] Example 1 - Plasmid DNA isolation and purification

[0200] The following is an exemplary procedure for preparing FGFR fusion plasmid DNA.

[0201] Equipment needed: centrifuge, capable of 1500 x g; microcentrifuge; pipette, positive displacement or exhaust; vortexer; nanodtop spectrophotometer; 37 °C shaker / incubator; and set at 37 ℃ oven.

[0202] Materials needed: frozen glycerol bacterial stock containing plasmid DNA; Kanamycin LB Agar Plates (Teknova #L1155); LB Broth (Life Technologies #10855-021); Kanamycin (Sigma #K0254); Plasmid Purification Kit (Qiagen # 12123); Pure Ethanol (Sigma Aldrich # E7023); Isopropanol (Sigma Aldrich #W292907); Nuclease-free water (without DEPC treatment) (from IDT or Ambion # AM9932); RNase barrier (filter) tips; RNase-free microtubes (1.5 to 2 mL VWR# 10011-724); serological pipettes; and 14ml round bottom tubes (VWR#352057).

[0203] To recover bacteria from the glycerol stock, use the tip of a sterile pipette to scrape frozen...

Embodiment 2

[0205] Example 2 - Generation of NRK cell lines

[0206] Expression vectors expressing each FGFR fusion were constructed. The expression vector was then transfected into normal rat kidney epithelial (NRK) cells. After transfection, stable cell lines were selected in media containing kanamycin. These cells were then grown, mRNA isolated and subjected to a FGFR fusion assay to confirm the presence of specific FGFR fusion mRNA.

Embodiment 3-F

[0207] Example 3-FGFR fusion cell line maintenance

[0208] The following protocol describes exemplary methods for culturing and maintaining NRK FGFR fusion overexpressing cell lines. Cell lines include, but are not limited to: NRK / FGFR3:TACC3v1, NRK / FGFR3:TACC3 v3, NRK / FGFR3:BAIAP2L1, NRK / FGFR2:BICC1, NRK / FGFR2:CASP7, NRK / FGFR2:CCDC6, NRK / FGFR2:AFF3, NRK / FGFR2:OFD1 and NRK / EMPTY VECTOR (plasmid control).

[0209] Required equipment: Biological safety cabinet, equipped with a vacuum suction system; CO 2 Incubator, set to 5% CO 2 and 37°C; a -80°C refrigerator; a liquid nitrogen tank; a water bath, set at 37°C; and a microscope.

[0210] Materials needed: Serological pipettes; Tissue culture flasks (T75 VWR # BD353136 and / or T150 VWR #15705-074); Tissue culture 0.2 μm filter units (Thermo Scientific #566-0020); Gell's Medium) Cell Culture Medium (Life Technologies, #11965-084); Fetal Bovine Serum (FBS), Certified Heat Inactivated (Life Technologies, #10082147); PenStrep A...

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Abstract

Disclosed herein are methods of identifying a cancer patient that will be responsive to treatment with a fibroblast growth factor receptor (FGFR) inhibitor and methods of treating cancer patients. The methods involve evaluating a biological sample from the patient for the presence of one or more FGFR mutants from a FGFR mutant gene panel. Kits and primers for identifying the presence of one or more FGFR mutant genes in a biological sample are also disclosed herein.

Description

[0001] Cross References to Related Applications [0002] This application claims priority to US Provisional Application No. 62 / 056,159, filed September 26, 2014, the disclosure of which is hereby incorporated by reference in its entirety. [0003] sequence listing [0004] This application contains a Sequence Listing that has been filed electronically in ASCII format, which is hereby incorporated by reference in its entirety. Said ASCII copy was created on August 6, 2015, named 103693.000782_SL.txt, and was 66,185 bytes in size. technical field [0005] Provided herein are methods of identifying cancer patients who will respond to treatment with fibroblast growth factor receptor inhibitors and methods of treating said cancer patients. Background technique [0006] Identifying genetic abnormalities can be used to select one or more appropriate treatments for cancer patients. It is also useful for cancer patients whose cancer type cannot be treated with the primary regimen...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6886C12Q2600/106C12Q2600/156A61K31/498A61P35/00
Inventor J.卡科拉S.J.普拉特罗
Owner JANSSEN PHARMA NV
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