PMID- 31601865
OWN - NLM
STAT- MEDLINE
DCOM- 20201111
LR  - 20231213
IS  - 2045-2322 (Electronic)
IS  - 2045-2322 (Linking)
VI  - 9
IP  - 1
DP  - 2019 Oct 10
TI  - Macrophage polarization impacts tunneling nanotube formation and intercellular 
      organelle trafficking.
PG  - 14529
LID - 10.1038/s41598-019-50971-x [doi]
LID - 14529
AB  - Tunneling nanotubes (TNTs) are cellular extensions enabling cytosol-to-cytosol 
      intercellular interaction between numerous cell types including macrophages. 
      Previous studies of hematopoietic stem and progenitor cell (HSPC) transplantation 
      for the lysosomal storage disorder cystinosis have shown that HSPC-derived 
      macrophages form TNTs to deliver cystinosin-bearing lysosomes to cystinotic 
      cells, leading to tissue preservation. Here, we explored if macrophage 
      polarization to either proinflammatory M1-like M(LPS/IFNgamma) or anti-inflammatory 
      M2-like M(IL-4/IL-10) affected TNT-like protrusion formation, intercellular 
      transport and, ultimately, the efficacy of cystinosis prevention. We designed new 
      automated image processing algorithms used to demonstrate that LPS/IFNgamma 
      polarization decreased bone marrow-derived macrophages (BMDMs) formation of 
      protrusions, some of which displayed characteristics of TNTs, including 
      cytoskeletal structure, 3D morphology and size. In contrast, co-culture of 
      macrophages with cystinotic fibroblasts yielded more frequent and larger 
      protrusions, as well as increased lysosomal and mitochondrial intercellular 
      trafficking to the diseased fibroblasts. Unexpectedly, we observed normal 
      protrusion formation and therapeutic efficacy following disruption of 
      anti-inflammatory IL-4/IL-10 polarization in vivo by transplantation of HSPCs 
      isolated from the Rac2(-/-) mouse model. Altogether, we developed unbiased image 
      quantification systems that probe mechanistic aspects of TNT formation and 
      function in vitro, while HSPC transplantation into cystinotic mice provides a 
      complex in vivo disease model. While the differences between polarization cell 
      culture and mouse models exemplify the oversimplicity of in vitro cytokine 
      treatment, they simultaneously demonstrate the utility of our co-culture model 
      which recapitulates the in vivo phenomenon of diseased cystinotic cells 
      stimulating thicker TNT formation and intercellular trafficking from macrophages. 
      Ultimately, we can use both approaches to expand the utility of TNT-like 
      protrusions as a delivery system for regenerative medicine.
FAU - Goodman, Spencer
AU  - Goodman S
AUID- ORCID: 0000-0001-5825-5055
AD  - Department of Pediatrics, Division of Genetics, University of California, San 
      Diego, La Jolla, California, USA.
FAU - Naphade, Swati
AU  - Naphade S
AD  - Department of Pediatrics, Division of Genetics, University of California, San 
      Diego, La Jolla, California, USA.
FAU - Khan, Meisha
AU  - Khan M
AD  - Department of Pediatrics, Division of Genetics, University of California, San 
      Diego, La Jolla, California, USA.
FAU - Sharma, Jay
AU  - Sharma J
AD  - Department of Pediatrics, Division of Genetics, University of California, San 
      Diego, La Jolla, California, USA.
FAU - Cherqui, Stephanie
AU  - Cherqui S
AUID- ORCID: 0000-0003-1240-5219
AD  - Department of Pediatrics, Division of Genetics, University of California, San 
      Diego, La Jolla, California, USA. scherqui@ucsd.edu.
LA  - eng
GR  - P30 NS047101/NS/NINDS NIH HHS/United States
GR  - R01 NS108965/NS/NINDS NIH HHS/United States
GR  - R01 DK090058/DK/NIDDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20191010
PL  - England
TA  - Sci Rep
JT  - Scientific reports
JID - 101563288
RN  - 0 (Cytokines)
RN  - 147336-22-9 (Green Fluorescent Proteins)
RN  - EC 3.6.5.2 (rac GTP-Binding Proteins)
SB  - IM
MH  - Animals
MH  - Apoptosis
MH  - Coculture Techniques
MH  - Cystinosis/metabolism
MH  - Cytokines/metabolism
MH  - Cytoskeleton/metabolism
MH  - Female
MH  - Fibroblasts/cytology
MH  - Green Fluorescent Proteins/metabolism
MH  - Hematopoietic Stem Cells/cytology
MH  - Inflammation
MH  - Lysosomes/metabolism
MH  - Macrophages/*cytology
MH  - Male
MH  - Mice
MH  - Mice, Inbred C57BL
MH  - Mice, Transgenic
MH  - Nanotubes/*chemistry
MH  - Organelles/*metabolism
MH  - Phenotype
MH  - Stem Cells/cytology
MH  - rac GTP-Binding Proteins/genetics
MH  - RAC2 GTP-Binding Protein
PMC - PMC6787037
COIS- Stephanie Cherqui is a cofounder, shareholder and a member of both the Scientific 
      Board and Board of Directors of GenStem Therapeutics Inc. Stephanie Cherqui 
      serves as a consultant for AVROBIO and receives compensation for these services. 
      Stephanie Cherqui also serves as a member of the Scientific Review Board and 
      Board of Trustees of the Cystinosis Research Foundation. The terms of this 
      arrangement have been reviewed and approved by the University of California San 
      Diego in accordance with its conflict of interest policies.
EDAT- 2019/10/12 06:00
MHDA- 2020/11/12 06:00
PMCR- 2019/10/10
CRDT- 2019/10/12 06:00
PHST- 2019/04/15 00:00 [received]
PHST- 2019/09/23 00:00 [accepted]
PHST- 2019/10/12 06:00 [entrez]
PHST- 2019/10/12 06:00 [pubmed]
PHST- 2020/11/12 06:00 [medline]
PHST- 2019/10/10 00:00 [pmc-release]
AID - 10.1038/s41598-019-50971-x [pii]
AID - 50971 [pii]
AID - 10.1038/s41598-019-50971-x [doi]
PST - epublish
SO  - Sci Rep. 2019 Oct 10;9(1):14529. doi: 10.1038/s41598-019-50971-x.