Dissecting hematopoietic and renal cell heterogeneity in adult zebrafish at single-cell resolution using RNA sequencing.

Qin Tang, Sowmya Iyer, Riadh Lobbardi, John C Moore, Huidong Chen, Caleb Lareau, Christine Hebert, McKenzie L Shaw, Cyril Neftel, Mario L Suva, Craig J Ceol, Andre Bernards, Martin Aryee, Luca Pinello, Iain A Drummond, David M Langenau
Author Information
  1. Qin Tang: Molecular Pathology Unit, Massachusetts General Hospital, Charlestown, MA. ORCID
  2. Sowmya Iyer: Molecular Pathology Unit, Massachusetts General Hospital, Charlestown, MA.
  3. Riadh Lobbardi: Molecular Pathology Unit, Massachusetts General Hospital, Charlestown, MA.
  4. John C Moore: Molecular Pathology Unit, Massachusetts General Hospital, Charlestown, MA. ORCID
  5. Huidong Chen: Department of Biostatistics, T.H. Chan School of Public Health, Harvard University, Boston, MA.
  6. Caleb Lareau: Center for Computational and Integrative Biology, Massachusetts General Hospital, Charlestown, MA. ORCID
  7. Christine Hebert: Molecular Pathology Unit, Massachusetts General Hospital, Charlestown, MA.
  8. McKenzie L Shaw: Molecular Pathology Unit, Massachusetts General Hospital, Charlestown, MA.
  9. Cyril Neftel: Molecular Pathology Unit, Massachusetts General Hospital, Charlestown, MA.
  10. Mario L Suva: Molecular Pathology Unit, Massachusetts General Hospital, Charlestown, MA.
  11. Craig J Ceol: Department of Molecular, Cell and Cancer Biology, University of Massachusetts Medical School, Worcester, MA. ORCID
  12. Andre Bernards: Center for Cancer Research, Massachusetts General Hospital, Charlestown, MA.
  13. Martin Aryee: Center for Computational and Integrative Biology, Massachusetts General Hospital, Charlestown, MA.
  14. Luca Pinello: Molecular Pathology Unit, Massachusetts General Hospital, Charlestown, MA. ORCID
  15. Iain A Drummond: Division of Nephrology, Department of Medicine, Massachusetts General Hospital, Boston, MA.
  16. David M Langenau: Molecular Pathology Unit, Massachusetts General Hospital, Charlestown, MA dlangenau@mgh.harvard.edu. ORCID

Abstract

Recent advances in single-cell, transcriptomic profiling have provided unprecedented access to investigate cell heterogeneity during tissue and organ development. In this study, we used massively parallel, single-cell RNA sequencing to define cell heterogeneity within the zebrafish kidney marrow, constructing a comprehensive molecular atlas of definitive hematopoiesis and functionally distinct renal cells found in adult zebrafish. Because our method analyzed blood and kidney cells in an unbiased manner, our approach was useful in characterizing immune-cell deficiencies within DNA-protein kinase catalytic subunit (), interleukin-2 receptor γ a (), and double-homozygous-mutant fish, identifying blood cell losses in T, B, and natural killer cells within specific genetic mutants. Our analysis also uncovered novel cell types, including two classes of natural killer immune cells, classically defined and erythroid-primed hematopoietic stem and progenitor cells, mucin-secreting kidney cells, and kidney stem/progenitor cells. In total, our work provides the first, comprehensive, single-cell, transcriptomic analysis of kidney and marrow cells in the adult zebrafish.

References

  1. Science. 2016 Jan 8;351(6269):aab2116 [PMID: 26541609]
  2. Cell Stem Cell. 2008 Feb 7;2(2):183-9 [PMID: 18371439]
  3. J Exp Med. 2016 Nov 14;213(12 ):2575-2589 [PMID: 27810924]
  4. J Exp Med. 2016 May 30;213(6):979-92 [PMID: 27139488]
  5. PLoS One. 2012;7(5):e37877 [PMID: 22655075]
  6. Curr Opin Genet Dev. 2000 Jun;10(3):252-6 [PMID: 10826982]
  7. Blood. 2002 Nov 1;100(9):3175-82 [PMID: 12384415]
  8. Dev Biol. 1997 Aug 15;188(2):189-204 [PMID: 9268568]
  9. Science. 1993 Dec 17;262(5141):1880-3 [PMID: 8266078]
  10. Cell Rep. 2016 Feb 2;14 (4):966-977 [PMID: 26804912]
  11. Cell Stem Cell. 2012 Jun 14;10(6):690-7 [PMID: 22704509]
  12. Genome Res. 2017 Mar;27(3):451-461 [PMID: 28087841]
  13. Nat Med. 2016 Feb;22(2):210-6 [PMID: 26752517]
  14. Nat Biotechnol. 2014 Apr;32(4):381-386 [PMID: 24658644]
  15. Blood. 2013 Aug 22;122(8):e1-11 [PMID: 23861249]
  16. Int Immunol. 1996 Jun;8(6):939-49 [PMID: 8671683]
  17. Nature. 2011 Feb 3;470(7332):95-100 [PMID: 21270795]
  18. Bioinformatics. 2013 Jan 1;29(1):15-21 [PMID: 23104886]
  19. Development. 1998 Dec;125(23):4655-67 [PMID: 9806915]
  20. Nat Protoc. 2017 Jan;12 (1):44-73 [PMID: 27929523]
  21. Blood. 2006 Dec 15;108(13):3976-8 [PMID: 16926288]
  22. J Am Soc Nephrol. 2015 Nov;26(11):2669-77 [PMID: 25817355]
  23. Proc Natl Acad Sci U S A. 2004 May 11;101(19):7369-74 [PMID: 15123839]
  24. J Leukoc Biol. 2006 Dec;80(6):1281-8 [PMID: 16963624]
  25. Nat Cell Biol. 2017 Apr;19(4):271-281 [PMID: 28319093]
  26. Cell. 1992 Mar 6;68(5):855-67 [PMID: 1547487]
  27. Cell. 2015 Jan 15;160(1-2):241-52 [PMID: 25594182]
  28. Nat Methods. 2013 Nov;10(11):1096-8 [PMID: 24056875]
  29. Blood. 2016 Aug 25;128(8):e20-31 [PMID: 27365425]
  30. Science. 1994 Mar 11;263(5152):1453-4 [PMID: 8128231]
  31. Nat Methods. 2017 Oct;14 (10 ):979-982 [PMID: 28825705]
  32. J Exp Med. 2017 Apr 3;214(4):1011-1027 [PMID: 28351983]
  33. Cell. 2015 May 21;161(5):1187-1201 [PMID: 26000487]
  34. Science. 1993 Dec 17;262(5141):1874-7 [PMID: 8266076]
  35. Blood. 1995 Jan 1;85(1):38-42 [PMID: 7803808]
  36. Science. 2017 Mar 31;355(6332): [PMID: 28360267]
  37. Dev Comp Immunol. 2015 Jul;51(1):148-59 [PMID: 25813149]
  38. Results Probl Cell Differ. 2017;60:55-75 [PMID: 28409342]
  39. Science. 2002 Jul 5;297(5578):99-102 [PMID: 12098699]
  40. Blood. 2011 Jan 27;117(4):e49-56 [PMID: 21084707]
  41. Oncogene. 2004 Sep 20;23(43):7233-46 [PMID: 15378083]
  42. Bioinformatics. 2014 Apr 1;30(7):923-30 [PMID: 24227677]
  43. Blood. 2005 Dec 1;106(12):3803-10 [PMID: 16099879]
  44. Blood. 1997 Mar 15;89(6):1968-77 [PMID: 9058718]
  45. Science. 2016 Apr 8;352(6282):189-96 [PMID: 27124452]
  46. Cell Stem Cell. 2008 Aug 7;3(2):169-81 [PMID: 18682239]
  47. Dev Biol. 2012 Aug 1;368(1):4-17 [PMID: 22664176]
  48. Genesis. 2001 Apr;29(4):156-62 [PMID: 11309848]
  49. Genome Biol. 2014;15(12):550 [PMID: 25516281]

Grants

  1. R01 AR063850/NIAMS NIH HHS
  2. R01 DK053093/NIDDK NIH HHS
  3. R24 OD016761/NIH HHS
  4. UH3 DK107372/NIDDK NIH HHS

MeSH Term

Animals
Animals, Genetically Modified
Cell Lineage
Gene Expression Profiling
Hematopoiesis, Extramedullary
Hematopoietic Stem Cells
Kidney
RNA
Sequence Analysis, RNA
Zebrafish

Chemicals

RNA

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