Gene expression data on >20 year-old, paired frozen and archival FFPE liver samples generated using targeted resequencing (TempO-Seq) and whole-genome RNA-sequencing methods. Samples were originally collected from male mice exposed to a reference chemical (dichloroacetic acid, DCA) at 0, 198, 313 and 427 mg/kg-day, (n=6/dose) by drinking water for 6 days. Portions of this dataset are inaccessible because: Including all of these files on ScienceHub would exceed the 1 Gb limit. They can be accessed through the following means: The data is stored on the L: Drive. Information on all of the files and file paths are indicated in the 20210923_Readme.xlsx file uploaded with the data. Format: Excel files, tab delimited files, csv files, and sequencing FASTQ files. This dataset is associated with the following publication: Cannizzo, M., C. Wood, S. Hester, and L. Wehmas. Case study: Targeted RNA-sequencing of aged formalin-fixed paraffin-embedded samples for understanding chemical mode of action. Toxicology Reports. Elsevier B.V., Amsterdam, NETHERLANDS, 9: 883-894, (2022).

Low RNA yield and quality limit use of formalin-fixed paraffin-embedded (FFPE) tissue samples for genomic analyses. In this study, we evaluated methods to demodify RNA highly fragmented and crosslinked by formalin fixation. Primary endpoints were RNA recovery, RNA-sequencing quality metrics, and target gene responses to a reference chemical (phenobarbital, PB). Frozen mouse liver samples from control and PB groups (n=6/group) were divided and preserved for 3 months as follows: frozen (FR); 70% ethanol (OH); 10% buffered formalin for 18 hours followed by ethanol (18F); and 10% buffered formalin (3F). Samples from OH, 18F, and 3F groups were processed to FFPE blocks and sectioned for RNA isolation. The latter group received no additional treatment (3F) or the following demodification protocols: short heated incubation with TAE buffer; overnight heated incubation with an organocatalyst using two different isolation kits; or overnight heated incubation without organocatalyst. TruSeq Stranded Total RNA libraries with Ribo-Zero were built and sequenced using the Illumina HiSeq platform. Extended incubation with or without organocatalyst increased RNA yield >3-fold and enhanced quality compared to 3F, as indicated by higher RNA integrity number (>1.5-fold) and fragment analysis values (>3.0-fold). Post-sequencing metrics showed reduced bias in gene coverage and deletion rates for all extended incubation groups. Following PB-induced differential gene expression analysis, all demodification groups showed increased overlap with FR in genes (73-83%) and pathways (91-94%) compared to 3F overlap with FR (60% and 63%, respectively). These results demonstrate simple changes in RNA isolation methods that can enhance genomic analyses of FFPE samples. This dataset is associated with the following publication: Wehmas, L., C. Wood, R. Gagne, A. Williams, C. Yauk, M. Gosink, D. Dalmas, R. Hao, R. O'Lone, and S. Hester. Demodifying RNA for Transcriptomic Analyses of Archival Formalin-Fixed Paraffin-Embedded Samples. TOXICOLOGICAL SCIENCES. Society of Toxicology, RESTON, VA, 162(2): 535-547, (2018).

Low RNA yield and quality limit use of formalin-fixed paraffin-embedded (FFPE) tissue samples for genomic analyses. In this study, we evaluated methods to demodify RNA highly fragmented and crosslinked by formalin fixation. Primary endpoints were RNA recovery, RNA-sequencing quality metrics, and target gene responses to a reference chemical (phenobarbital, PB). Frozen mouse liver samples from control and PB groups (n=6/group) were divided and preserved for 3 months as follows: frozen (FR); 70% ethanol (OH); 10% buffered formalin for 18 hours followed by ethanol (18F); and 10% buffered formalin (3F). Samples from OH, 18F, and 3F groups were processed to FFPE blocks and sectioned for RNA isolation. The latter group received no additional treatment (3F) or the following demodification protocols: short heated incubation with TAE buffer; overnight heated incubation with an organocatalyst using two different isolation kits; or overnight heated incubation without organocatalyst. TruSeq Stranded Total RNA libraries with Ribo-Zero were built and sequenced using the Illumina HiSeq platform. Extended incubation with or without organocatalyst increased RNA yield >3-fold and enhanced quality compared to 3F, as indicated by higher RNA integrity number (>1.5-fold) and fragment analysis values (>3.0-fold). Post-sequencing metrics showed reduced bias in gene coverage and deletion rates for all extended incubation groups. Following PB-induced differential gene expression analysis, all demodification groups showed increased overlap with FR in genes (73-83%) and pathways (91-94%) compared to 3F overlap with FR (60% and 63%, respectively). These results demonstrate simple changes in RNA isolation methods that can enhance genomic analyses of FFPE samples. This dataset is associated with the following publication: Wehmas, L., C. Wood, R. Gagne, A. Williams, C. Yauk, M. Gosink, D. Dalmas, R. Hao, R. O'Lone, and S. Hester. Demodifying RNA for Transcriptomic Analyses of Archival Formalin-Fixed Paraffin-Embedded Samples. TOXICOLOGICAL SCIENCES. Society of Toxicology, RESTON, VA, 162(2): 535-547, (2018).

The primary goal of this study was to investigate the effects of whether organocatalyst (ORGΔ) treatment improves DNA-sequencing data by overcoming formalin-induced artifacts from clinical formalin-fixed paraffin-embedded (FFPE) samples. We isolated RNA and DNA ± ORGΔ from paired FFPE and frozen human renal and ovarian carcinoma specimens collected as part of the National Cancer Institute Biospecimen Pre-analytical Variables program. Tumor types were microscopically confirmed from adjacent tissue sections. Following extraction, DNA was fragmented and sequence differences were compared between frozen and FFPE sample pairs. Treatment with ORGΔ improved concurrent SNP calls in FFPE DNA compared to non-ORGΔ FFPE samples and enhanced confidence in SNP calls for all FFPE DNA samples, beyond that of matched frozen samples. In general, the concordant SNPs identified in paired frozen and FFPE DNA samples agreed for both genotype and homozygosity vs. heterozygosity of calls regardless of ORGΔ treatment. The increased confidence in ORGΔ FFPE DNA variant calls relative to the matched frozen DNA suggests a novel application of this method. With further optimization, this method may improve quality of DNA-sequencing data in frozen as well as FFPE tissue samples. These findings could have important implications for studies using FFPE samples, especially if adequate matched controls are not available. This dataset is associated with the following publication: Wehmas, L., C. Wood, P. Guan, M. Gosink, and S. Hester. Organocatalyst treatment improves variant calling and mutant detection in archival clinical samples. Scientific Data. Springer Nature, New York, NY, 12: 6509, (2022).

se of archival resources has been limited to date by inconsistent methods for genomic profiling of degraded RNA from formalin-fixed paraffin-embedded (FFPE) samples. RNA-seq offers a novel way to address this problem. In this study we evaluated transcriptomic dose responses using RNA-seq in paired FFPE and frozen (FROZ) samples from two archival studies in mice, one recent (<2 years old) and the other older (>20 years old). Experimental treatments included di(2-ethylhexyl)phthalate (DEHP) and dichloroacetic acid (DCA) for the <2 and >20 year-old studies, respectively. Total RNA was ribodepleted and sequenced using the Illumina HiSeq platform. In the recent study, FFPE samples showed high concordance in total reads (98% vs FROZ), fold-change values of differentially expressed genes (DEGs) (R2 = 0.99), highly enriched target pathways (90% overlap with FROZ), and benchmark dose estimates for preselected target genes (-2% overall vs FROZ). In contrast, RNA-seq data from older FFPE samples had lower total reads (70% vs FROZ) and poor concordance in global DEGs and pathways. Despite a 99% loss of counts, dose responses were still evident for target genes in FFPE samples and positively correlated with paired FROZ samples. These findings highlight potential variability in the quality of RNA-seq data from FFPE samples. More recent FFPE samples were highly similar to FROZ samples in sequencing quality metrics, DEG profiles, and dose-response parameters, while further methods development is needed for older or lower-quality FFPE samples. This work should help broaden the use of archival resources in both chemical safety and translational science. This dataset is associated with the following publication: Hester, S., V. Bhat, B. Chorley, G. Carswell, W. Jones, L. Wehmas, and C. Wood. Dose-Response Analysis of RNA-Seq Profiles in Archival Formalin-Fixed Paraffin-Embedded (FFPE) Samples.. TOXICOLOGICAL SCIENCES. Society of Toxicology, 154(2): 202-213, (2016).

RNA sequencing data from paired: (1) fresh-frozen, (2) direct-fixed in formalin for 18 hours, (3) frozen then formalin-fixed, or (4) frozen then ethanol-fixed and paraffin-embedded (n=6/group/condition) tissue samples from the livers of mice exposed to 600 ppm phenobarbital vs. vehicle control for 7 days in drinking water. RNA sequencing data from paired fresh frozen and 18 hr. formalin-fixed paraffin-embedded liver tissue from mice administered 1500, 3000, and 6000 ppm DEHP or vehicle control for 7 days by dietary ingestion; or RNA sequencing data from paired fresh frozen, 18 hr. formalin fixed paraffin embedded, or 3 wk. formalin fixed paraffin embedded liver tissue from mice administered 8 mg/kg/d Furan or vehicle control for 3 weeks by gavage. This dataset is associated with the following publication: Wehmas, L., S. Hester, and C.E. Wood. Direct Formalin Fixation Induces Widespread Transcriptomic Effects in Archival Tissue Samples. Scientific Reports. Nature Publishing Group, London, UK, 10: 14497, (2020).

File contains raw data collected on body and organ weights of mice given PFBS and body burden of the chemical after intervals of several hours, as well as expression of liver candidate genes for nuclear receptors at 24 h post-treatment. This dataset is associated with the following publication: Lau, C., J. Rumpler, K. Das, C. Wood, J. Schmid, M. Strynar, and J. Wambaugh. Pharmacokinetic profile of Perfluorobutane Sulfonate and activation of hepatic nuclear receptor target genes in mice. TOXICOLOGY. Elsevier Science Ltd, New York, NY, USA, 441: 152522, (2020).

File contains raw data collected on body and organ weights of mice given PFBS and body burden of the chemical after intervals of several hours, as well as expression of liver candidate genes for nuclear receptors at 24 h post-treatment. This dataset is associated with the following publication: Lau, C., J. Rumpler, K. Das, C. Wood, J. Schmid, M. Strynar, and J. Wambaugh. Pharmacokinetic profile of Perfluorobutane Sulfonate and activation of hepatic nuclear receptor target genes in mice. TOXICOLOGY. Elsevier Science Ltd, New York, NY, USA, 441: 152522, (2020).

This file contains a link for Gene Expression Omnibus and the GSE designations for the publicly available gene expression data used in the study and reflected in Figures 6 and 7 for the Das et al., 2016 paper. This dataset is associated with the following publication: Das, K., C. Wood, M. Lin, A.A. Starkov, C. Lau, K.B. Wallace, C. Corton, and B. Abbott. Perfluoroalky acids-induced liver steatosis: Effects on genes controlling lipid homeostasis. TOXICOLOGY. Elsevier Science Ltd, New York, NY, USA, 378: 32-52, (2017).

Data set 1 consists of the experimental data for the Wild Type and PPAR KO animal study and includes data used to prepare Figures 1-4 and Table 1 of the Das et al, 2016 paper. This dataset is associated with the following publication: Das, K., C. Wood, M. Lin, A.A. Starkov, C. Lau, K.B. Wallace, C. Corton, and B. Abbott. Perfluoroalky acids-induced liver steatosis: Effects on genes controlling lipid homeostasis. TOXICOLOGY. Elsevier Science Ltd, New York, NY, USA, 378: 32-52, (2017).