Evolutionary switch in expression of key markers between mouse and human leads to mis-assignment of cell types in developing adrenal medulla

We would like to clarify the key aspects of ongoing discussion (Bedoya-Reina and Schlisio, 2021Bedoya-Reina O.C. Schlisio S. Chromaffin cells with sympathoblast signature: too similar keep apart?.Cancer Cell. 2021; 39: 134-135Abstract Full Text PDF PubMed Scopus (5) Google Scholar; Dong et al., 2020Dong R. Yang Zhan Y. Lai H.D. Ye C.J. Yao X.Y. Luo W.Q. Cheng X.M. Miao J.J. Wang J.F. al.Single-cell characterization malignant phenotypes developmental trajectories adrenal neuroblastoma.Cancer 2020; 38: 716-733Abstract (49) Kildisiute 2021bKildisiute G. Young M.D. Behjati Pitfalls applying mouse markers human medullary cells.Cancer 132-133Abstract 2021Yang W. Li K. J. Response Kildsiute al. Bedoya-Reina Schlisio.Cancer 136-137Abstract (3) Scholar) regarding recent study by (Dong Scholar), in which authors performed single-cell transcriptomics analysis developing medulla, followed comparison heterogeneous neuroblastoma cell populations. are grateful for publishing their valuable data, we believe that communal work will interpretation data context embryos pathological tissues. When assigning clusters, annotated population based on expression a literature-derived marker gene: CARTPT Scholar). In contrast, other groups Jansky 2021Jansky Sharma A.K. Körber V. Quintero A. Toprak U.H. Wecht E.M. Gartlgruber M. Greco Chomsky E. Grünewald T.G.P. transcriptomic analyses provide insights into origins neuroblastoma.Nat. Genet. https://doi.org/10.1038/s41588-021-00806-1Crossref (33) Kameneva 2021Kameneva P. Artemov A.V. Kastriti M.E. Faure L. Olsen T.K. Otte Erickson Semsch B. Andersson E.R. Ratz identifies multiple lineages potential implications origin.Nat. https://doi.org/10.1038/s41588-021-00818-xCrossref (29) assigned same cluster chromaffin fate, multi-gene “annotation transfer” analysis. While Cartpt is well-validated as sympathoblasts (Furlan 2017Furlan Dyachuk Calvo-Enrique Abdo H. Hadjab Chontorotzea T. Akkuratova N. Usoskin D. Kamenev al.Multipotent peripheral glial generate neuroendocrine medulla.Science. 2017; 357: eaal3753Crossref (146) our (Kameneva well validations shown below, revealed that, humans, instead expressed predominantly cells. This striking inconsistency different organisms has led incorrect assignments clusters original manuscript Humans differ from mice, it natural expect there be non-conservative situations number genes. Here, reanalyzed The datasets were merged analyzed standard Seurat (Stuart 2019Stuart Butler Hoffman Hafemeister C. Papalexi Mauck 3rd, W.M. Hao Stoeckius Smibert Satija Comprehensive integration data.Cell. 2019; 177: 1888-1902Abstract (3392) (Figures S1A–S1C). Setting aside non-conserved gene, clear known does not agree annotation suggested Specifically, sympathetic expresses set clear-cut including PNMT, PENK, shows higher levels CHGB S1B, S1D, S1E). opposite cluster, interpreted cells, PRPH, ELAVL4 clearly represents neurons response criticism raised teams co-authors demonstrated results immunohistochemistry (Yang showing tight intra-adrenal negative CART. Thus, they concluded these represent CART− However, aggregations forming ganglia-like structures tissue immunopositive neurofilament heavy chain, HUC/D (proteins encoded ELAVL3/4 genes) S2A S2B) study, CART immunoreactivity was never observed maturing but evident identified TH+HUC/D− across several stages S2C S2D). only rare immature neuroblasts show some (Figure S2C, yellow arrows) at early (week 8), those transient likely become according data. type found lineage-tracing originate Schwann precursors Consistently, connect via bridge both humans clustering generated us 2021aKildisiute Kholosy Roberts Elmentaite van Hooff S.R. Pacyna C.N. Khabirova Piapi Thevanesan al.Tumor normal mRNA comparisons reveal pan-neuroblastoma cancer cell.Sci. Adv. 7: eabd3311Crossref S2). Likewise, extra-adrenal CART+ week 8 became during later development S2E). meantime, systematically increases through all investigated stages. cannot used reliable approach assign or fates embryonic tumor populations, labels much stronger longer compared neurons. Overall, few genes turned out sensitive fluctuating evolutionary conservation gene and, thus, required tighter experimental major studied organism. interesting observation also highlights need rely broader signatures cross-species orthologous types. Download .pdf (.15 MB) Help pdf files Document S1. Figures S1 S2 Cells Sympathoblast Signature: Too Similar Keep Apart?Bedoya-Reina al.Cancer CellDecember 31, 2020In BriefDong (2020) built an atlas neural crest derivatives trace neuroblastoma. murine anlagen designated resemble mouse. Full-Text Open ArchiveSingle-Cell Characterization Malignant Phenotypes Developmental Trajectories Adrenal NeuroblastomaDong CellSeptember 17, detailed phenotypic landscape (NB) using RNA sequencing proliferation/differentiation status highly associated clinical heterogeneity NB. ArchiveResponse SchlisioYang BriefWe appreciate attention comments publication 2020). letters interest communities many bioinformaticians. ArchivePitfalls Applying Mouse Markers Human Medullary CellsKildisiute BriefIn this letter, re-analyze fetal demonstrate pitfalls relying annotate unbiased label transfer methods imply had suggested. Archive