Multi-omic cell free DNA profiles of cerebral spinal fluid from lung cancer metastasis to the brain.
The 7 matches · 1 of them tie a paragraph to a whole file, not to given lines: a weak match, whose lines are not tinted
- [1] § Results › Specific 5mC methylation patterns present in the CSF cfDNA of brain metastasis ↔ CSF_cfDNA_5mC_5hmC_analysis.R, lines 54–104 · score 0.75 · IL1R2, RAD54B, CCNH, CD74, ZKSCAN1, MID1
- [2] § Results › Comparing the methylation of brain metastasis CSF cfDNA to NSCLC primary tumor DNA ↔ CSF_cfDNA_5mC_5hmC_analysis.R, lines 54–104 · score 0.68 · ATAD3A, COL6A6, RAD54B, MIF, MMP13, overlapped
- [3] § Results › Decreased intragenic hydroxymethylation in the CSF cfDNA of brain metastasis ↔ CSF_cfDNA_5mC_5hmC_analysis.R, lines 106–146 · score 0.68 · IL21R, ADCY9, HDAC9, KCNQ1, MTDH, PBX1
- [4] § Results › Decreased intragenic hydroxymethylation in the CSF cfDNA of brain metastasis ↔ cfDNA_enrichr_analysis.R, lines 52–91 · score 0.57 · PPI Hub Proteins, Transcription Factor, hmCG, Enrichr, bar
- [5] § Materials and methods › Methylation and hydroxymethylation analysis of CSF cfDNA ↔ CSF_cfDNA_5mC_5hmC_analysis.R, lines 1–52 · score 0.56 · GENCODE v38, intersecting, FDR, hydroxymethylation, protein, promoter
- [6] § Results › Features of CSF cfDNA level among patients with brain metastasis ↔ CSF_cfDNA_sequencing_coverage.R, the whole file · a weak match · score 0.54 · sequencing coverage, CSF volumes, CSF samples, DNA, cancer
- [7] § Results › CSF cfDNA profiles reveal longitudinal changes from clinical events ↔ CSF_plasma_cfDNA_size_comparision.R, lines 189–229 · score 0.54 · dt_ratio, md_ratio, mt_ratio, nucleosome ratios, dinucleosome, trinucleosome
Paper
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The authors' code
R · 253 lines · 9.5 KB · MIT · 4 matches
- # CSF_cfDNA_5mC_5hmC_analysis.R
- # this file is meant to be used inside Rstudio
- # this script takes in promoter methylation (5mC) & intragenic hydroxymethylation (5hmC) % per CpG site with fdr-corrected q values, groups by gene level
- # and plots the promoter 5mC% or intragenic 5hmC% per gene (volcano plot) or per sample (point plot) for both cancer and control
- # this file was generated using bedtools intersect with GENCODE v38:
- # bedtools intersect -a gencode.v38.genes.sorted.bed -b P1.5mC.sorted.bedgraph -wa -wb -sorted | awk '{print($0 '\tP1')}' > P1.5mC.sorted.genes.intersect.bed
- # P1.5mC.sorted.bedgraph is a bedgraph of the modified_bases.5mC.bed from dorado, filtered on column 5 > 0, and extracting only the chrom, start, end, and percent methylation columns
- # multiple intersected bed files can be joined using cat, and fed into this script.
- library(data.table)
- library(dplyr)
- library(ggplot2)
- library(ggrepl)
- library(tidyverse)
- # plot differential prmoter 5mCG% and intragenic 5hmCG% between cancer and control CSF of all genes in volcano plot
- # read in merged bed files
- # promoter 5mCG
- hyper_pro_5mc <- fread("/mnt/ix1/Projects/M085_221117_CSF_cfDNA_methylation/03_statistical_test/04_volcano_plot/02_samples/gene_summary/hyper.2kbpromoter_summary_5mCG.txt")
- hypo_pro_5mc <- fread("/mnt/ix1/Projects/M085_221117_CSF_cfDNA_methylation/03_statistical_test/04_volcano_plot/02_samples/gene_summary/hypo.2kbpromoter_summary_5mCG.txt")
- # intragenic 5hmCG
- hyper_5hmc <- fread("/mnt/ix1/Projects/M085_221117_CSF_cfDNA_methylation/03_statistical_test/04_volcano_plot/02_samples/gene_summary/hyper.genebody_summary_5hmCG.txt")
- hypo_5hmc <- fread("/mnt/ix1/Projects/M085_221117_CSF_cfDNA_methylation/03_statistical_test/04_volcano_plot/02_samples/gene_summary/hypo.genebody_summary_5hmCG.txt")
- # gene analysis
- # diff_methyl = 5mCG% (cancer ) - 5mCG% (control) per gene
- # promoter 5mCG
- # use gencodev38 gene names to filter protein_coding genes
- gencode <- fread("/mnt/ix1/Projects/M086_221117_TGP_cfDNA_methylation/annotation/gencode.v38.genes_summary.txt")
- gencode_proteincoding <- gencode %>% filter(V5=='protein_coding')
- protein_coding_genes <- unique(gencode_proteincoding$V6)
- hyper_pro_5mc_mean <- hyper_pro_5mc %>%
- select(c('V5', 'V6', 'V10', 'V11')) %>%
- rename(gene = V5, gene_id = V6,
- diff_methyl = V10, qval = V11) %>%
- filter(gene %in% protein_coding_genes)
- hypo_pro_5mc_mean <- hypo_pro_5mc %>%
- select(c('V5', 'V6', 'V10', 'V11')) %>%
- rename(gene = V5, gene_id = V6,
- diff_methyl = V10, qval = V11) %>%
- filter(gene %in% protein_coding_genes)
- df_pro_5mc <- rbind(hyper_pro_5mc_mean, hypo_pro_5mc_mean)
- # add legend to the plot
- df_pro_5mc <- df_pro_5mc %>%
- mutate(group = case_when(
- qval < 0.01 & diff_methyl > 0 ~ "hypermethylated in cancer",
- qval < 0.01 & diff_methyl < 0 ~ "hypomethylated in cancer",
- TRUE ~ "Background"))
- # add reported marker labels on the plot
- df_marker_pro_5mc <- df_pro_5mc %>%
- filter(gene %in% c('ATAD3A','CCNH','CD74','DYRK2','IL1R2',
- 'LIPH','MID1','MIF','MMP13','RAD54B',
- 'SETDB1','ZKSCAN1','COL6A6',
- 'EPB41L1','IL6R','PER3')) %>%
- filter(qval<0.01) %>% group_by(gene, diff_methyl,qval) %>%
- summarize(n = n())
- # plot the volcano plot
- p1_2 <- ggplot(data=df_pro_5mc, aes(x=diff_methyl, y=-log10(1e-5+qval))) +
- geom_point(aes(color=group),size=6, alpha=0.6) +
- scale_color_manual(name ='type',
- values = c("hypermethylated in cancer" = "#CC79A7",
- "hypomethylated in cancer" = "#009E73")) +
- geom_text_repel(data = df_marker_pro_5mc,
- aes(x= diff_methyl, y=-log10(1e-5+qval), label = gene),
- min.segment.length = 0,
- max.overlaps=Inf,
- size = 8 ) +
- labs(x='CpG promoter 5mCG difference per gene (%)') +
- theme_classic(30) +
- theme(axis.text = element_text(face = 'bold'),
- axis.title = element_text(face = 'bold'))
- p1_2
- # intragenic 5hmCG
- hyper_5hmc_mean <- hyper_5hmc %>%
- select(c('V5', 'V6', 'V10', 'V11')) %>%
- rename(gene_type = V5, gene = V6,
- diff_methyl = V10, qval = V11) %>%
- filter(gene_type == 'protein_coding')
- hypo_5hmc_mean <- hypo_5hmc %>%
- select(c('V5', 'V6', 'V10', 'V11')) %>%
- rename(gene_type = V5, gene = V6,
- diff_methyl = V10, qval = V11) %>%
- filter(gene_type == 'protein_coding')
- df_5hmc <- rbind(hyper_5hmc_mean, hypo_5hmc_mean)
- # add legend to the plot
- df_5hmc <- df_5hmc %>%
- mutate(group = case_when(
- qval < 0.01 & diff_methyl > 0 ~ "hyper-hydroxymethylated in cancer",
- qval < 0.01 & diff_methyl < 0 ~ "hypo-hydroxymethylated in cancer",
- TRUE ~ "Background"))
- # add reported marker labels on the plot
- df_marker_5hmc <- df_5hmc %>%
- filter(gene %in% c('ADCY9','HDAC9','IL21R','KCNQ1',
- 'MTDH','PBX1','SASH1','TPST1','TWIST2')) %>%
- filter(qval<0.01) %>% group_by(gene, diff_methyl,qval) %>%
- summarize(n = n())
- # plot the volcano plot
- p1_2 <- ggplot(data=df_5hmc, aes(x=diff_methyl, y=-log10(1e-6 + qval))) +
- geom_point(aes(color=group),size=6, alpha=0.6) +
- scale_color_manual(name ='type', values = c("hyper-hydroxymethylated in cancer" = "#CC79A7",
- "hypo-hydroxymethylated in cancer" = "#009E73")) +
- geom_text_repel(data = df_marker_5hmc,
- aes(x = diff_methyl, y = -log10(1e-6 + qval), label = gene),
- min.segment.length = 0,
- max.overlaps = Inf,
- size = 8) +
- labs(x='CpG intragenic 5hmCG difference per gene (%)') +
- theme_classic(30) +
- theme(axis.text = element_text(face = 'bold'),
- axis.title = element_text(face = 'bold'))
- p1_2
- ggsave("5mCG_promoter_volcano.png", plot = p1_1, dpi = 300, units = "in", height = 12, width = 20)
- ggsave("5hmCG_intragenic_volcano.png", plot = p1_2, dpi = 300, units = "in", height = 12, width = 20)
- # plot differential prmoter 5mCG% and intragenic 5hmCG% of marker genes per sample
- # these marker genes were defined as sig genes (q<0.01) between cancer vs. control CSF samples, and reported in the literature as biomarkers for NSCLC
- # e.g. a sig gene that was hypomethylated in our data and reported as an upregulated marker/overexpressed in NSCLC patients
- # promoter 5mCG
- p2_1 <- ggplot(marker_5mc_pro, aes(x= gene, y= mean_methyl, color = type)) +
- geom_jitter(size=6, alpha=0.6, width = 0.3, height = 2) +
- #scale_y_continuous(limits = c(0,100), breaks = c(0,50,100)) +
- geom_vline(xintercept = c(1.5,2.5,3.5,4.5,5.5,6.5,7.5,8.5,9.5,
- 10.5,11.5,12.5,13.5,14.5,15.5),
- linetype="dotted",size=0.5) +
- labs(x = "Gene" ,y ="CpG promoter 5mCG
- per sample (%)") +
- theme_classic(25) +
- theme(axis.text = element_text(face = "bold"),
- axis.title = element_text(face = "bold"),
- axis.text.x = element_text(angle = -45, hjust = 0.05))
- p2_1
- # intragenic 5hmCG
- p2_2 <- ggplot(marker_5hmc, aes(x= gene, y= mean_methyl, color = type)) +
- geom_jitter(size=6, alpha=0.6, width = 0.3, height = 2) +
- #scale_y_continuous(limits = c(0,100), breaks = c(0,50,100)) +
- geom_vline(xintercept = c(1.5,2.5,3.5,4.5,5.5,6.5,7.5,8.5),
- linetype="dotted",size=0.5) +
- labs(x = "Gene" ,y ="CpG intragenic 5hmCG
- per sample (%)") +
- theme_classic(30) +
- theme(axis.text = element_text(face = "bold"),
- axis.title = element_text(face = "bold"),
- axis.text.x = element_text(angle = -45, hjust = 0.05))
- p2_2
- ggsave("5mCG_promoter_marker_plot.png", plot = p2_1, dpi = 300, units = "in", height = 8, width = 20)
- ggsave("5hmCG_intragenic_marker_plot.png", plot = p2_2, dpi = 300, units = "in", height = 10, width = 16)
- # plot average genome-wide 5hmCG% between cancer and control CSF samples
- # read in the merged bedmethyl files for 5hmCG
- # these are bedgraphs containing 5hmC% before intersecting with GENCODE
- cpg_5hmc <- fread("/mnt/ix1/Projects/M085_221117_CSF_cfDNA_methylation/00_sample/mp0007_sample_summary_sup_methyl/sorted_bedgraph/5hmCG/merged.genomewide.5hmCG.txt")
- cpg_5hmc <- cpg_5hmc %>% mutate(type = ifelse(grepl('control',V5),'Healthy','Cancer'))
- # calculate average genome-wide 5hmCG%
- avg_5hmc <- cpg_5hmc %>% group_by(V5,type) %>%
- summarise(mean_5hmCG = mean(V4)) %>%
- mutate(m = signif(mean_5hmCG,2))
- # plot
- p3_1 <- ggplot(avg_5hmc, aes(x= type, y= mean_5hmCG, fill = type)) +
- geom_boxplot(width=0.7) +
- labs(x = NULL, y ="Average genome-wide
- 5hmCG (%)") +
- #scale_y_continuous(limits = c(0.8,9), breaks = c(1,3,5,7,9)) +
- theme_classic(25) +
- theme(axis.text = element_text(face = "bold"),
- axis.title = element_text(face = "bold"))
- p3_1
- p3_2 <- ggplot(avg_5hmc, aes(x= m, fill = type)) +
- geom_bar(width=0.1, color ='black') +
- labs(x = "Average genome-wide 5hmCG (%)", y ="Number of samples", fill = "Type") +
- #scale_x_continuous(limits = c(54,72), breaks = c(54,60,66,72)) +
- #scale_y_continuous(limits = c(0,2), breaks = c(0,1,2)) +
- facet_wrap(~ type, ncol = 1) +
- theme_classic(40) +
- theme(strip.text = element_text(size = 40),
- axis.text = element_text(face = "bold"),
- axis.title = element_text(face = "bold"),
- panel.margin = unit(0, "lines"))
- p3_2
- # caculate p value for the box plot
- p_5hmc <- avg_5hmc %>% summarise(p=t.test(mean_5hmCG[type=='Healthy'], mean_5hmCG[type=='Cancer'])$p.value)
- ggsave("genomewide_avg_5hmCG_boxplot.png", plot = p3_1, dpi = 300, units = "in", height = 6, width = 8)
- ggsave('genomewide_avg_5hmCG_barplot.png', plot = p3_2, dpi = 300, units = 'in', height = 10, width = 20)
CSF_cfDNA_5mC_5hmC_analysis.R at commit e393d61, under MIT · at the source
Overview
- Division of Oncology, Department of Medicine, Stanford University, School of Medicine, Stanford, CA 94305, United States
- Department of Neurosurgery, Stanford University, Stanford, CA 94305, United States
- Department of Electrical Engineering, Stanford University, Stanford, CA 94305, United States
Abstract
Cerebrospinal fluid (CSF) is a liquid biological medium immersing the brain and spinal canal. For patients with metastatic cancer from the lung, tumor cells release their cell-free DNA (cfDNA) into the CSF. We applied a nanopore single-molecule sequencing approach to analyze the fragmentation, methylation, and hydroxymethylation patterns present in CSF-derived cell-free DNA from patients with metastatic lung cancer to the brain. We compared the cancer cfDNA finding to non-cancer healthy controls and their CSF cell-free DNA. Among cancer patients, we observed enriched mono-nucleosome levels and significantly higher mono-/
Reproduced under the paper's license (CC BY-NC), from the paper cited above.
Repositories
Its files are read in the Code ↔ Paper reader above, with 7 matches between paragraphs and lines of code.
Tianqi-Kiki/nanopore_CSF_cfDNA
e393d614799ee1a94bcbb550de6d920429c50c69, 24 July 2025Availability: 1 check, the latest on 27 September 2026: the link answers
- 27 September 2026: the link answers
7 files
- CSF_cfDNA_5mC_5hmC_analy
sis.R , R, 253 lines, 4 matches - CSF_cfDNA_sequencing_cov
erage.R , R, 92 lines, 1 match - CSF_cfDNA_size_analysis.
R , R, 150 lines - CSF_plasma_cfDNA_size_co
mparision.R , R, 337 lines, 1 match - cfDNA_enrichr_analysis.R
, R, 209 lines, 1 match - LICENSE, License, 21 lines
- README.md, Text, 46 lines
Zenodo 20173042
Availability: 1 check, the latest on 27 September 2026: the link answers (HTTP 200)
- 27 September 2026: the link answers (HTTP 200)
7 files
- CSF_cfDNA_5mC_5hmC_analy
sis.R , R, 253 lines - CSF_cfDNA_sequencing_cov
erage.R , R, 92 lines - CSF_cfDNA_size_analysis.
R , R, 150 lines - CSF_plasma_cfDNA_size_co
mparision.R , R, 337 lines - cfDNA_enrichr_analysis.R
, R, 209 lines - LICENSE, License, 21 lines
- README.md, Text, 46 lines
The paper's code and data availability statement is in the Data section.
Tracing map
Proposed by the machine: these links were found in the paper and verified at the source, without human review. The map will receive a Zenodo DOI once one of the paper's authors has validated it with their ORCID.
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Its JSON (tracing-map.json) is deposited on Zenodo with its DOI once the map is validated.
Data
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Data availability
Sequence-aligned BAM files and associated methylation calls are deposited in NCBI’s dbGAP under accession phs003794.v1.p1 (https://
Reproduced under the paper's license (CC BY-NC), from the paper cited above.
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Version 1, 27 September 2026: the first record
Recorded: type, language, journal, volume, issue, pages, dates, 7 authors, 15 MeSH terms, 2 funders, 85 references.
Cite
This paper
Chen, T., Bai, X., Burnside, G., Trinh, T. T. H., Gephart, M. H., Ji, H. P., & Lau, B. T. (2026). Multi-omic cell free DNA profiles of cerebral spinal fluid from lung cancer metastasis to the brain. NAR cancer, 8(3), zcag021. https://
BibTeX
@article{chen2026multi,
author = {Chen, Tianqi and Bai, Xiangqi and Burnside, Georgiana and Trinh, Thy Trang Hoang and Gephart, Melanie Hayden and Ji, Hanlee P and Lau, Billy T},
title = {{Multi-omic cell free DNA profiles of cerebral spinal fluid from lung cancer metastasis to the brain}},
journal = {NAR cancer},
year = {2026},
month = aug,
volume = {8},
number = {3},
pages = {zcag021},
publisher = {Oxford University Press},
issn = {2632-8674},
doi = {10.1093/
url = {https://
pmid = {42676708},
pmcid = {PMC13527165}
}
RIS
TY - JOUR
AU - Chen, Tianqi
AU - Bai, Xiangqi
AU - Burnside, Georgiana
AU - Trinh, Thy Trang Hoang
AU - Gephart, Melanie Hayden
AU - Ji, Hanlee P
AU - Lau, Billy T
TI - Multi-omic cell free DNA profiles of cerebral spinal fluid from lung cancer metastasis to the brain
T2 - NAR cancer
J2 - NAR Cancer
PY - 2026
DA - 2026/
VL - 8
IS - 3
SP - zcag021
SN - 2632-8674
PB - Oxford University Press
DO - 10.1093/
UR - https://
LA - en
ER -
CSL-JSON
{
"id": "10.1093/
"type": "article-journal",
"title": "Multi-omic cell free DNA profiles of cerebral spinal fluid from lung cancer metastasis to the brain",
"container-title": "NAR cancer",
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"family": "Chen",
"given": "Tianqi"
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{
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"given": "Georgiana"
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{
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}
],
"container-title-short":
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"issue": "3",
"page": "zcag021",
"DOI": "10.1093/
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"PMCID": "PMC13527165",
"ISSN": "2632-8674",
"publisher": "Oxford University Press",
"URL": "https://
"language": "en",
"issued": {
"date-parts": [
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