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Next Generation Sequencing Facilitates Quantitative Analysis of MM cell line Transcriptomes

ABSTRACT: Next Generation Sequencing Facilitates Quantitative Analysis of MM cell line Transcriptomes

PROVIDER: PRJNA756303 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR15530267_1.fastq.gz	Fastqsanger.gz
	SRR15530267_2.fastq.gz	Fastqsanger.gz
	SRR15530268.fastq.gz	Fastqsanger.gz
	SRR15530268_1.fastq.gz	Fastqsanger.gz
	SRR15530268_2.fastq.gz	Fastqsanger.gz

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Next Generation Sequencing Facilitates Quantitative Analysis of MM cell line Transcriptomes

Project description:We discovered that the activated-leukocyte-cell-adhesion-molecule (ALCAM) regulated myelomagenesis. ALCAM was a negative regulator of myeloma clonogenicity. To determine what signalling pathways are affected by ALCAM in MM cells, we established CTR-KD and ALCAM-KD RPMI8226 MM cell lines. Hoechst33342 staining was performed on CTR-KD and ALCAM-KD RPMI8226 MM cell. Then flow cytometry was used to separate MP and SP cells of the two types of cells. Total RNA of each sample was extracted using TRIzol Reagent (Invitrogen). Next generation sequencing library preparations were constructed using NEBNext® Ultra™ RNA Library Prep Kit for Illumina (Vazyme, Cat No. NR603, China).RNA-seq was performed by GENEWIZ (Suzhou, China) on an Illumina Hiseq platform.

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Next Generation Sequencing Facilitates Quantitative Analysis of Transcriptomes of MM cells after M2 Treatment

Project description:Targeted immunotherapies based on monoclonal antibodies have continued to show clinical activities in patients with multiple myeloma (MM). A novel antibody drug conjugate targeting BCMA armed with a DNA cross-linking pyrrolobenzodiazepine PBD dimer tesirine M2, is currently under clinical development for MM. M2 has demonstrated potent cytotoxicity against MM cells in bulk as well as quiescent tumor-initiating cells in in vitro and in vivo preclinical studies. We here further investigate its effects on transcriptional changes in MM cells prior to apoptosis and identify molecular events regulating its potential immunomodulation to further upregulate its efficacy. RNA sequencing followed by gene set enrichment analysis in MM cells treated with M2 vs control vehicle at sub-lethal concentrations reveal that M2 can induce interferon-regulated genes dependent on DNA damage-initiated signaling pathways, providing new molecular insight into its mechanisms of action in MM in the bone marrow microenvironment.

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