Immuno-oncologic profiling by stage-dependent transcriptome and proteome analyses of spontaneously regressing canine cutaneous histiocytoma

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Bioinformatics and Genomics

Main article text

 

Introduction

Materials and Methods

Selection of tissue samples

Preparation of tissue samples

RNA extraction from FFPE samples and RNA quality control

3′ RNA sequencing (RNA-seq)

Functional enrichment analysis

RNA hybridization using nCounter

In situ hybridization

Counting of CD80 and CD86-positive tumor cells

MALDI-MSI

Protein identification by electrospray ionization tandem mass spectrometry

Statistical analysis

Results

Only minor differences between the compared groups on the mRNA level

Only minor differences between compared groups on protein level

Components of MHC I and MHC II are expressed in tumor cell-enriched groups of CCH on the mRNA and protein levels

Higher expression of co-stimulatory molecules than co-inhibitory molecules in CCH tumor cell-enriched sample groups on the mRNA level

Increasing expression of CD80 and constantly high expression of CD86 in tumor cells in the time course of CCH regression

Lower expression of CD86 but not CD80 in HS compared to CCH tumor cells

Discussion

Conclusions

Supplemental Information

mRNA expression levels of round cell tumor markers according to Paździor-Czapula et al., 2015 from canine cutaneous histiocytoma groups 1, 2a, 2b and 3.

Box-and-whisker plots display values as maximum, minimum, median, lower and upper quartiles (n = 5 or 6) of normalized counts as detected by QunatSeq 3′ analysis. The mean baseMean of all detected RNAs (132, dashed line) is provided as a basis for comparison to the number of respective round cell tumor marker counts.

DOI: 10.7717/peerj.18444/supp-1

mRNA expression patterns of CD80, CD86.

Ornithine decarboxylase antizyme 1 (OAZ1) and dihydrodipicolinate reductase (DapB) in tumor cells of canine cutaneous histiocytoma (CCH) and canine histiocytic sarcoma (HS), as well as lymphocytes, epidermis, blood vessels and apocrine glands of tumor adjacent tissue of CCH. OAZ1 served as control for RNA accessibility. DapB served as a negative control. I. situ hybridization with fast red (chromogen, red) and Mayer’s hematoxylin (blue) counterstain. Magnification: 600×.

DOI: 10.7717/peerj.18444/supp-2

RNA quality number and DV200 of samples used for QuantSeq 3′ and nCounter Canine IO Panel analyses.

DOI: 10.7717/peerj.18444/supp-3

Differential expressed genes of QuantSeq 3′ analysis.

DOI: 10.7717/peerj.18444/supp-4

List of gene sets investigated using the nCounter Canine IO Panel.

DOI: 10.7717/peerj.18444/supp-5

Abundantly expressed peptides from MALDI-MSI.

DOI: 10.7717/peerj.18444/supp-6

Results log2FC 0.5.

DOI: 10.7717/peerj.18444/supp-7

Additional Information and Declarations

Competing Interests

The authors declare that they have no competing interests.

Author Contributions

Alina K. Loriani Fard conceived and designed the experiments, performed the experiments, analyzed the data, prepared figures and/or tables, authored or reviewed drafts of the article, and approved the final draft.

Alexander Haake analyzed the data, authored or reviewed drafts of the article, and approved the final draft.

Vladimir Jovanovic analyzed the data, prepared figures and/or tables, authored or reviewed drafts of the article, and approved the final draft.

Sandro Andreotti analyzed the data, prepared figures and/or tables, authored or reviewed drafts of the article, and approved the final draft.

Michael Hummel conceived and designed the experiments, authored or reviewed drafts of the article, and approved the final draft.

Benjamin-Florian Hempel conceived and designed the experiments, analyzed the data, authored or reviewed drafts of the article, and approved the final draft.

Achim D. Gruber conceived and designed the experiments, analyzed the data, authored or reviewed drafts of the article, and approved the final draft.

Data Availability

The following information was supplied regarding data availability:

QuantSeq 3′ data and nCounter data are available at the Gene Expression Omnibus (GEO) Database: GSE261387 and GSE261395, respectively.

Mass spectrometry proteomics (.raw) and imaging (.imzML) data are available at the ProteomeXchange Consortium via the MassIVE partner repository under project name “Immuno-Oncologic Profiling by Stage-Dependent Transcriptome and Proteome Analyses of Spontaneously Regressing Canine Cutaneous Histiocytoma”: PXD050403.

Funding

This study was supported by “Förderlinie TEAMS Mittel zur Förderung der Vernetzung zwischen Wissenschaftler/-innen der Freien Universität Berlin” (TEAMS funding line Funds to promote networking between Scientists at the Freie Universität Berlin). There was no additional external funding received for this study. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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