CoChat
Sign Up
Cited by 36Mesenchymal stem cell researchCRISPR and Genetic EngineeringPluripotent Stem Cells Research

Targeting ectodysplasin promotor by CRISPR/dCas9-effector effectively induces the reprogramming of human bone marrow-derived mesenchymal stem cells into sweat gland-like cells

Open access
Sun, Sujing, Xiao, Jun, Huo, Jiahui, Geng, Zhijun, Ma, Kui, Sun, Xiaoyan, Fu, Xiaobing·2018·Stem Cell Research & Therapy
DOI 10.1186/s13287-017-0758-0PMC PMC5766979
Open this paper in CoChat — free accountOpen PDF

Abstract

BACKGROUND: Patients with a deep burn injury are characterized by losing the function of perspiration and being unable to regenerate the sweat glands. Because of their easy accession, multipotency, and lower immunogenicity, bone marrow-derived mesenchymal stem cells (BM-MSCs) represent as an ideal biological source for cell therapy. The aim of this study was to identify whether targeting the promotor of ectodysplasin (EDA) by CRISPR/dCas9-effector (dCas9-E) could induce the BM-MSCs to differentiate into sweat gland-like cells (SGCs). METHODS: Activation of EDA transcription in BM-MSCs was attained by transfection of naive BM-MSCs with the lenti-CRISPR/dCas9-effector and single-guide RNAs (sgRNAs). The impact of dCas9-E BM-MSCs on the formation of SGCs and repair of burn injury was identified and evaluated both in vitro and in a mouse model. RESULTS: After transfection with sgRNA-guided dCas9-E, the BM-MSCs acquired significantly higher transcription and expression of EDA by doxycycline (Dox) induction. Intriguingly, the specific markers (CEA, CK7, CK14, and CK19) of sweat glands were also positive in the transfected BM-MSCs, suggesting that EDA plays a critical role in promoting BM-MSC differentiation into sweat glands. Furthermore, when the dCas9-E BM-MSCs with Dox induction were implanted into a wound in a laboratory animal model, iodine-starch perspiration tests revealed that the treated paws were positive for perspiration, while the paws treated with saline showed a negative manifestation. For the regulatory mechanism, the expression of downstream genes of NF-κB (Shh and cyclin D1) was also enhanced accordingly. CONCLUSIONS: These results suggest that EDA is a pivotal factor for sweat gland regeneration from BM-MSCs and may also offer a new approach for destroyed sweat glands and extensive deep burns.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1186/s13287-017-0758-0

Cites

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • Recent studies have demonstrated the potential conversion of autologous somatic cells, such as epidermal progenitors or mesenchymal stem cells, into SwG lineages through reprogramming manipulations.

    “Recent studies have demonstrated the potential conversion of autologous somatic cells, such as the epidermal progenitors[ 3 ] or mesenchymal stem cells,[ 4 ] into SwG lineages through reprogramming manipulations.”

    From Sweat Gland Organoids Originating from Reprogrammed Epidermal Keratinocytes Functionally Recapitulated Damaged Skin (2021) · cites this paper as [4]

    The Results section reports that reprogrammed human bone marrow-derived mesenchymal stem cells acquired sweat gland markers, supporting the mesenchymal-stem-cell portion attributed to this citation. The Methods section says the cells were commercially purchased; it does not establish that the converted cells were autologous. The epidermal-progenitor portion is attached to a different citation. (LLM, conf=0.86)

    Experimental — accuracy not yet measured

  • A previous study demonstrated the conversion of bone marrow-derived mesenchymal stem cells into SwG cells via forced EDA gene expression.

    “Our previous study demonstrated the conversion of bone marrow‐derived mesenchymal stem cells into SwG cells via forced EDA gene expression,[ 4 ]”

    From Sweat Gland Organoids Originating from Reprogrammed Epidermal Keratinocytes Functionally Recapitulated Damaged Skin (2021) · cites this paper as [4]

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

The citation lists haven’t been fetched yet

References 33 · Cited by 36

Research works printed but not linked: 31

  1. [1]Xu Y Huang S Ma K Fu X Han W Sheng Z Promising new potential for mesenchymal stem cells derived from human umbilical cord Wharton's jelly: sweat gland cell-like differentiative capacity J Tissue Eng Regen Med 2012 6 645 54 10.1002/term.468 21916019 · DOI
  2. [2]Fu X Fang L Li X Cheng B Sheng Z Enhanced wound-healing quality with bone marrow mesenchymal stem cells autografting after skin injury Wound Repair Regen 2006 14 325 35 10.1111/j.1743-6109.2006.00128.x 16808812 · DOI
  3. [3]Xu Y Hong Y Xu M Ma K Fu X Zhang M Wang G Role of keratinocyte growth factor in the differentiation of sweat gland-like cells from human umbilical cord-derived mesenchymal stem cells Stem Cells Transl Med 2016 5 106 16 10.5966/sctm.2015-0081 26574554 PMC4704873 · DOI
  4. [4]Bianco P Riminucci M Gronthos S Robey PG Bone marrow stromal stem cells: nature, biology, and potential applications Stem Cells 2001 19 180 92 10.1634/stemcells.19-3-180 11359943 · DOI
  5. [5]Sheng Z Fu X Cai S Lei Y Sun T Bai X Chen M Regeneration of functional sweat gland-like structures by transplanted differentiated bone marrow mesenchymal stem cells Wound Repair Regen 2009 17 427 35 10.1111/j.1524-475X.2009.00474.x 19660052 · DOI
  6. [6]Xu Y Huang S Fu X Autologous transplantation of bone marrow-derived mesenchymal stem cells: a promising therapeutic strategy for prevention of skin-graft contraction Clin Exp Dermatol 2012 37 497 500 10.1111/j.1365-2230.2011.04260.x 22300217 · DOI
  7. [7]Kearns NA Genga RM Enuameh MS Garber M Wolfe SA Maehr R Cas9 effector-mediated regulation of transcription and differentiation in human pluripotent stem cells Development 2014 141 219 23 10.1242/dev.103341 24346702 PMC3865759 · DOI
  8. [9]Smith C, Ye Z, Cheng L. Genome editing in human pluripotent stem cells. Cold Spring Harbor protocols. 2016;2016:pdb top086819. 27037079
  9. [10]Ding Q Regan SN Xia Y Oostrom LA Cowan CA Musunuru K Enhanced efficiency of human pluripotent stem cell genome editing through replacing TALENs with CRISPRs Cell Stem Cell 2013 12 393 4 10.1016/j.stem.2013.03.006 23561441 PMC3925309 · DOI
  10. [11]Mali P Yang L Esvelt KM Aach J Guell M DiCarlo JE Norville JE Church GM RNA-guided human genome engineering via Cas9 Science 2013 339 823 6 10.1126/science.1232033 23287722 PMC3712628 · DOI · Paper page
  11. [12]Werner S Keratinocyte growth factor: a unique player in epithelial repair processes Cytokine Growth Factor Rev 1998 9 153 65 10.1016/S1359-6101(98)00010-0 9754709 · DOI
  12. [13]Barrientos S Stojadinovic O Golinko MS Brem H Tomic-Canic M Growth factors and cytokines in wound healing Wound Repair Regen 2008 16 585 601 10.1111/j.1524-475X.2008.00410.x 19128254 · DOI · Paper page
  13. [14]Kunisada M Cui CY Piao Y Ko MS Schlessinger D Requirement for Shh and Fox family genes at different stages in sweat gland development Hum Mol Genet 2009 18 1769 78 10.1093/hmg/ddp089 19270025 PMC2671986 · DOI
  14. [15]Zhao Z Xu M Wu M Ma K Sun M Tian X Zhang C Fu X Direct reprogramming of human fibroblasts into sweat gland-like cells Cell Cycle 2015 14 3498 505 10.1080/15384101.2015.1093707 26566868 PMC4825576 · DOI
  15. [16]Cui CY Kunisada M Childress V Michel M Schlessinger D Shh is required for Tabby hair follicle development Cell Cycle 2011 10 3379 86 10.4161/cc.10.19.17669 21926481 PMC3233628 · DOI
  16. [17]Schmidt-Ullrich R Tobin DJ Lenhard D Schneider P Paus R Scheidereit C NF-kappaB transmits Eda A1/EdaR signalling to activate Shh and cyclin D1 expression, and controls post-initiation hair placode down growth Development 2006 133 1045 57 10.1242/dev.02278 16481354 · DOI · Paper page
  17. [18]Srivastava AK Durmowicz MC Hartung AJ Hudson J Ouzts LV Donovan DM Cui CY Schlessinger D Ectodysplasin-A1 is sufficient to rescue both hair growth and sweat glands in Tabby mice Hum Mol Genet 2001 10 2973 81 10.1093/hmg/10.26.2973 11751679 · DOI
  18. [19]Ra JC Shin IS Kim SH Kang SK Kang BC Lee HY Kim YJ Jo JY Yoon EJ Choi HJ Kwon E Safety of intravenous infusion of human adipose tissue-derived mesenchymal stem cells in animals and humans Stem Cells Dev 2011 20 1297 308 10.1089/scd.2010.0466 21303266 · DOI
  19. [20]Pispa J Thesleff I Mechanisms of ectodermal organogenesis Dev Biol 2003 262 195 205 10.1016/S0012-1606(03)00325-7 14550785 · DOI · Paper page
  20. [21]Cui CY Schlessinger D EDA signaling and skin appendage development Cell Cycle 2006 5 2477 83 10.4161/cc.5.21.3403 17102627 PMC2860309 · DOI
  21. [22]Perez-Pinera P Kocak DD Vockley CM Adler AF Kabadi AM Polstein LR Thakore PI Glass KA Ousterout DG Leong KW Guilak F Crawford GE Reddy TE Gersbach CA RNA-guided gene activation by CRISPR-Cas9-based transcription factors Nat Methods 2013 10 973 6 10.1038/nmeth.2600 23892895 PMC3911785 · DOI · Paper page
  22. [23]Gao Y Li M Zhang X Bai T Chi G Liu JY Li Y Isolation, culture and phenotypic characterization of human sweat gland epithelial cells Int J Mol Med 2014 34 997 1003 10.3892/ijmm.2014.1851 25187692 PMC4152140 · DOI
  23. [24]Li HH Zhou G Fu XB Zhang L Antigen expression of human eccrine sweat glands J Cutan Pathol 2009 36 318 24 10.1111/j.1600-0560.2008.01020.x 19032382 · DOI
  24. [25]Hammarstrom S The carcinoembryonic antigen (CEA) family: structures, suggested functions and expression in normal and malignant tissues Semin Cancer Biol 1999 9 67 81 10.1006/scbi.1998.0119 10202129 · DOI
  25. [26]Tao R Han Y Chai J Li D Sun T Isolation, culture, and verification of human sweat gland epithelial cells Cytotechnology 2010 62 489 95 10.1007/s10616-010-9303-z 20853024 PMC2995140 · DOI
  26. [27]Lamouille S Xu J Derynck R Molecular mechanisms of epithelial-mesenchymal transition Nat Rev Mol Cell Biol 2014 15 178 96 10.1038/nrm3758 24556840 PMC4240281 · DOI · Paper page
  27. [28]Hanson SE Bentz ML Hematti P Mesenchymal stem cell therapy for nonhealing cutaneous wounds Plast Reconstr Surg 2010 125 510 6 10.1097/PRS.0b013e3181c722bb 20124836 PMC4076140 · DOI
  28. [29]Pispa J Pummila M Barker PA Thesleff I Mikkola ML Edar and Troy signalling pathways act redundantly to regulate initiation of hair follicle development Hum Mol Genet 2008 17 3380 91 10.1093/hmg/ddn232 18689798 · DOI
  29. [30]Zhang Y Tomann P Andl T Gallant NM Huelsken J Jerchow B Birchmeier W Paus R Piccolo S Mikkola ML Morrisey EE Overbeek PA Scheidereit C Millar SE Schmidt-Ullrich R Reciprocal requirements for EDA/EDAR/NF-kappaB and Wnt/beta-catenin signaling pathways in hair follicle induction Dev Cell 2009 17 49 61 10.1016/j.devcel.2009.05.011 19619491 PMC2859042 · DOI
  30. [31]Newton K French DM Yan M Frantz GD Dixit VM Myodegeneration in EDA-A2 transgenic mice is prevented by XEDAR deficiency Mol Cell Biol 2004 24 1608 13 10.1128/MCB.24.4.1608-1613.2004 14749376 PMC344191 · DOI
  31. [32]Cho SW Kwak S Woolley TE Lee MJ Kim EJ Baker RE Kim HJ Shin JS Tickle C Maini PK Jung HS Interactions between Shh, Sostdc1 and Wnt signaling and a new feedback loop for spatial patterning of the teeth Development 2011 138 1807 16 10.1242/dev.056051 21447550 · DOI

Authors
Sun, S., Xiao, J., Huo, J., Geng, Z., Ma, K., Sun, X., & Fu, X.
Affiliations
  • Chinese PLA General Hospital (CN)
  • Air Force General Hospital PLA (CN)
Corresponding author
  • Sun, Xiaoyan
  • Fu, Xiaobing
ORCID
Venue
Stem Cell Research & Therapy
Publisher
Springer Science and Business Media LLC
Published
2018-01-12
Volume
9
Issue
1
Pages
8-8
ISSN
1757-6512 (ISSN-L)
DOI
10.1186/s13287-017-0758-0
PMC
PMC5766979
OpenAlex
W2783301241
Cited by
36
References
33
Citations by year
2025: 7 · 2024: 2 · 2023: 6 · 2022: 4 · 2021: 6 · 2020: 4 · 2019: 6 · 2018: 1
FWCI
2.41
Type
journal article
Language
en
Open access
Licence
cc-by
Locations
  • Stem Cell Research & Therapy (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
National Natural Science Foundation of China (81421064, 81372067)
Topics
  • Health Sciences › Medicine › Genetics › Mesenchymal stem cell research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
Keywords
ectodysplasin, bone marrow-derived mesenchymal stem cells, sweat gland regeneration, transcriptional activation, NF-κB signaling, Sonic hedgehog, cyclin D1, cell therapy, sweating
MeSH terms
Cellular Reprogramming Techniques, Gene Editing, RNA, Guide, CRISPR-Cas Systems, Animals, Bone Marrow Cells, Burns, Cell Differentiation, Cell Line, Disease Models, Animal, Doxycycline, Humans, Male, Mice, Inbred BALB C, Mice, Nude, Promoter Regions, Genetic, Sweat Glands, RNA, Guide, Kinetoplastida, Cyclin D1, Mesenchymal Stem Cell Transplantation, Mice, Ectodysplasins, Hedgehog Proteins, HEK293 Cells, Mesenchymal Stem Cells, CRISPR-Cas Systems, Cell- and Tissue-Based Therapy

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review