Biomarkers and methods for individualized treatment of small cell lung cancer using KDM1A inhibitors

A technology of KDM1A, treatment method, applied in the direction of biochemical equipment and methods, microbial determination/inspection, etc., can solve the problem that the sensitivity is not SCLC cells and so on

Pending Publication Date: 2022-04-12
奥莱松基因组股份有限公司
View PDF84 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, published data suggest that while certain SCLCs are highly sensitive to KDM1A inhibition, sensitivity to KDM1A inhibition is not a universal feature of SCLC cells

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Biomarkers and methods for individualized treatment of small cell lung cancer using KDM1A inhibitors
  • Biomarkers and methods for individualized treatment of small cell lung cancer using KDM1A inhibitors
  • Biomarkers and methods for individualized treatment of small cell lung cancer using KDM1A inhibitors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0143] Example 1: Sorting of KDM1A Inhibitor Sensitive and Resistant Cell Lines

[0144] For biomarker identification, seven SCLC cell lines were assessed for their response to KDM1A inhibitor treatment based on the results of viability assays performed 4 or 10 days after treatment with the KDM1A inhibitor ORY-1001 (as described elsewhere herein). Classification. For the 4-day viability assay, SCLC cell lines were seeded in 384-well plates in a final volume of 40 μL in the optimized medium supplemented with ORY-1001 recommended by the supplier of each cell line (maximum concentration: 50 μM; tested for 18 serial 1:2 dilutions). At 37°C, 5% CO 2 - After 4 days of incubation in a controlled atmosphere, use Assay Cell viability was assessed according to the manufacturer's protocol (Promega). EC50 values ​​were calculated using Microsoft Excel software for untreated cells (100% growth) and without Reagents (100% growth inhibition) were standardized. Viability quantificatio...

Embodiment 2

[0148] Example 2 Identification of ASCL1 and SOX2 as biomarkers of responsiveness to KDM1A inhibitors

[0149] To identify genes that can differentiate SCLC cell lines that are sensitive to KDM1A inhibitor therapy from those that are resistant, four KDM1Ai-sensitive, one KDM1Ai-partially sensitive, and two KDM1Ai-resistant SCLC cell lines were subjected to RNASeq analysis. Details on their responsiveness and classification to KDM1Ai are provided in Example 1 above.

[0150] Cell pellets for gene expression analysis

[0151] Cells were grown continuously in flasks for 6 days. On the last day of the assay, cells were collected in Falcon tubes, counted, and then centrifuged at 1200 rpm for 4 minutes. Discard the supernatant, suspend the cells in 1 mL of PBS and transfer to an eppendorf sample, centrifuge at 3000 rpm for 5 min in an eppendorf centrifuge at 4 °C. Finally, the supernatant was discarded and the pellet was frozen at -80°C.

[0152] RNA sequencing

[0153] To...

Embodiment 3

[0163] Example 3 Verification of ASCL1 and SOX2 using qRT-PCR

[0164] The same panel of SCLC cell lines described in Examples 1 and 2 (including two additional SCLC cell lines, one identified as sensitive to KDM1Ai treatment (DMS53) and the other identified as sensitive to KDM1Ai treatment) were then analyzed by Taqman qRT-PCR. KDM1Ai Treatment Partial Sensitivity (NCIH526)) The biomarkers of responsiveness to KDM1A inhibitors identified in Example 2, ASCL1 and SOX2, were validated.

[0165] Gene expression analysis by qRT-PCR

[0166] Total RNA was extracted using the RNeasy Mini kit, and cDNA was obtained using the High Capacity RNA-to-cDNA Master Mix (ThermoFisher Scientific #4390779) according to standard procedures. qRT-PCR was performed using a LightCycler 480 Probes Master (PNT-L-034; Roche #04887301001) and using a predesigned and preoptimized TaqMan gene expression assay from ThermoFisher Scientific. qRT-PCR was performed in triplicate using a Lightcycler 480 Ins...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

Disclosed are biomarkers and methods for predicting the responsiveness of a patient suffering from small cell lung cancer (SCLC) to treatment with a KDM1A inhibitor, and methods for treating a subgroup of SCLC patients identified using the methods.

Description

technical field [0001] The present invention relates to biomarkers and methods for personalized treatment of small cell lung cancer (SCLC) using KDM1A inhibitors. The present invention provides methods of identifying SCLC patients who may benefit from KDM1A inhibitor treatment and methods of treating such patients with KDM1A inhibitors. Background technique [0002] Lysine-specific demethylase 1 (LSD1, also known as KDM1A) is a histone-modifying enzyme responsible for the demethylation of dimethyl histone 3 lysine 4 (H3K4me2) (Shi et al. , Cell2004). In several human cancers, KDM1A overexpression has been associated with disease progression and poorer prognosis, and its inhibition has been shown to reduce cancer cell growth, migration and invasion. Therefore, KDM1A has been considered as a target of interest for the development of new drugs to treat cancer, and several KDM1A inhibitors are currently in clinical trials in oncology. [0003] In particular, KDM1A inhibitors ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/106C12Q2600/158
Inventor F·西赛里N·萨基洛托S·鲁纳迪
Owner 奥莱松基因组股份有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap