NBDC Research ID: hum0394.v2

 

SUMMARY

Aims: We perform spatial transcriptome analysis and other omics analyses, such as genome/epigenome sequencing and multiplexed immunofluorescence, using fresh-frozen and FFPE specimens of non-small cell lung cancers. We try to develop a novel stratification method for lung cancer patients by considering intratumor heterogeneity and conditions of complicated microenvironments of cancer tissues. We aim to characterize omics statuses of cancer cells and their surrounding stromal cells and immune cells, such as cancer-associated fibroblasts and tumor-infiltrating lymphocytes, at a spatial level and evaluate tumor cell-microenvironment relationships at a molecular level.

Methods: Spatial transcriptome sequencing Visium, high-performance in situ gene expression mapping Xenium, Multiplex Fluorescence Immunostaining, Whole Genome Sequencing and Targeted Long-Read Methylation Sequencing (t-nanoEM)

Participants/Materials:

     [Visium] 16 tumor sections from 8 non-small cell lung cancer patients

     [Xenium] Five tumor sections from 5 non-small cell lung cancer patients

     [WGS] 1 tumor section and 1 non-tumor section from 1 non-small cell lung cancer patient

     [t-nanoEM] Five microdissected tumor regions from 1 non-small cell lung cancer patient

 

Dataset IDType of DataCriteriaRelease Date
JGAS000613

NGS (Visium Spatial Gene Expression), Histological image

Xenium In Situ Gene Expression

Multiplex Fluorescence Immunostaining

Controlled-access (Type I) 2024/04/08
JGAS000757 NGS (WGS, t-nanoEM) Controlled-access (Type I) 2024/11/20

*Release Note

*Data users need to apply an application for Using NBDC Human Data to reach the Controlled-access Data. Learn more

 

MOLECULAR DATA

Visium Spatial Gene Expression, Histological image

Participants/Materials

non-small cell lung cancer (ICD10: C349): 8 cases

      surgical section: 16 samples

Targets Visium Spatial Gene Expression
Target Loci for Capture Methods -
Platform Illumina [NovaSeq 6000]
Library Source RNAs extracted from surgical sections (fresh-frozen/FFPE)
Cell Lines -
Library Construction (kit name) Visium Spatial Gene Expression Reagent Kits
Fragmentation Methods -
Spot Type Paired-end
Read Length (without Barcodes, Adaptors, Primers, and Linkers)

fresh-frozen: 118 bp

FFPE: 78 bp

Software spaceranger-1.2.1, spaceranger-1.3.0
QC -
Japanese Genotype-phenotype Archive Dataset ID JGAD000742
Total Data Volume 479.7 GB (fastq, png, json, csv, tsv, mtx, cloupe, html)
Comments (Policies) NBDC policy

 

Xenium In Situ Gene Expression

Participants/Materials

non-small cell lung cancer (ICD10: C349): 5 cases

      surgical section: 5 samples

Targets Xenium In Situ Gene Expression
Target Loci for Capture Methods 302 genes (Human Lung panel: 202 genes, custom: 100 genes)
Platform 10x Genomics [Xenium Analyzer]
Source surgical sections (fresh-frozen/FFPE)
Cell Lines -
Reagents (Panel, Kit, Version)

Xenium Slides & Sample Prep Reagents

Xenium Decoding Reagents

Xenium Decoding Consumables

Software xenium-1.1.0.2, xenium-1.2.0.3
Japanese Genotype-phenotype Archive Dataset ID JGAD000742
Total Data Volume 479.7 GB (csv, h5, html, json, mtx, tiff, parquet, tsv, xenium, zarr)
Comments (Policies) NBDC policy

 

Multiplex Fluorescence Immunostaining

Participants/Materials

lung adenocarcinoma (ICD10: C349): 3 cases

          tumor tissues: 3 samples

Targets Multiplex Fluorescence Immunostaining
Target Loci for Capture Methods cycle/probe information
Platform Akoya Biosciences [PhenoCycler]
Library Source tumor tissues of lung adenocarcinomas
Cell Lines -
Cell Treatment Method (kit name) Staining Kit for PhenoCycler
Japanese Genotype-phenotype Archive Dataset ID JGAD000742
Total Data Volume 479.7 GB (qptiff)
Comments (Policies) NBDC policy

 

WGS

Participants/Materials

non-small cell lung cancer (ICD10: C349): 1 case

          tumor tissue: 1 sample (Illumina, Nanopore)

          non-tumor tissue: 1 sample (Illumina)

Targets WGS
Target Loci for Capture Methods -
Platform

Illumina [NovaSeq 6000]

Nanopore [PromethION]

Library Source DNAs extracted from tumor tissue and non-tumor tissue
Cell Lines -
Library Construction (kit name)

Illumina: TruSeq Nano DNA Library Prep Kit, NEBNext Ultra II DNA Library Prep Kit and NEBNext Multiplex Oligos for Illumina

Nanopore: Ligation Sequencing Kit V14 (SQK-LSK114)

Fragmentation Methods

Illumina: Ultrasonic fragmentation (Covaris)

Nanopore: -

Spot Type

Illumina: Paired-end

Nanopore: Single-end

Read Length (without Barcodes, Adaptors, Primers, and Linkers)

Illumina: 300 bp

Nanopore: 7,500 bp on average

Mapping Methods

Illumina: BWA-MEM

Nanopore: Guppy

Mapping Quality -
Reference Genome Sequence hg38
Coverage (Depth) Illumina: x44 (non-tumor tissue), x82 (tumor tissue)
Japanese Genotype-phenotype Archive Dataset ID JGAD000898
Total Data Volume 616.6 GB (fastq, bam)
Comments (Policies) NBDC policy

 

t-nanoEM

Participants/Materials

non-small cell lung cancer (ICD10: C349): 1 case

          five microdissected tumor regions: 1 sample

Targets t-nanoEM
Target Loci for Capture Methods Twist Alliance Pan-cancer Methylation Panel - 1.5 MB
Platform Nanopore [PromethION]
Source DNA extracted from tumor tissue sample
Cell Lines -
Library Construction (kit name)

NEBNext Enzymatic Methyl-seq Kit

The five libraries were pooled and subjected to target enrichment using the Twist Standard Hyb and Wash Kit v2 with a pan-cancer panel.

Fragmentation Methods g-TUBE (Covaris)
Spot Type Single-end
Read Length (without Barcodes, Adaptors, Primers, and Linkers) 4,800 bp on average
Japanese Genotype-phenotype Archive Dataset ID JGAD000898
Total Data Volume 616.6 GB (fastq)
Comments (Policies) NBDC policy

 

DATA PROVIDER

Principal Investigator: Ayako Suzuki

Affiliation: Laboratory of Medical Omics Data Analysis, Department of Computational Biology and Medical Sciences, Graduate School of Frontier Sciences, The University of Tokyo

Project / Group Name: -

Funds / Grants (Research Project Number):

NameTitleProject Number
KAKENHI Grant-in-Aid for Early-Career Scientists Development of spatial genome and transcriptome analytical methods in local regions of lung cancer tissues 21K15566

 

PUBLICATIONS

TitleDOIDataset ID
1
2

 

USRES (Controlled-access Data)

Principal InvestigatorAffiliationCountry/RegionResearch TitleData in Use (Dataset ID)Period of Data Use