Intracellular signaling mechanisms associated with CD47 modified surfaces.
We have previously established that recombinant CD47 can ameliorate the inflammatory response to synthetic polymeric surfaces. Here, we begin to profile, at the transcriptional, translational and cell signaling level, the inflammatory cell response when blood interacts with CD47 modified polyvinyl chloride (PVC) (CD47-PVC). We used qPCR arrays to compare transcriptional changes between human whole blood exposed to CD47-PVC or PVC. Transcription of IL1F5, IL1F10, IL17F, CCL3, CCL8, CCL28, CXCL12, and CXCL13 was upregulated in blood exposed to PVC, compared to CD47-PVC. The increase in CCL3 and CCL8 transcription correlated with an increase in the chemokines' presence in the plasma. Exposure of blood to CD47-PVC resulted in an increase, compared to PVC, in transcription of CCL2, CCL4, CCL20, CXCL1, TGFβ3, GDF3, GDF10, CD40LG, and TNFSF10. CD47-PVC exposure resulted in an increase of the following matrix metalloproteinase related genes: MMP1, MMP7, MMP13, and MMP16. Phosflow cytometry, and assays examining transcription factor binding, cell attachment, and genome-wide chromatin association indicated that members of the JAK-STAT signaling pathway, particularly JAK2 and STAT5, mediate inflammatory cell interactions with CD47-PVC. Our data demonstrate that differential molecular responses to CD47 involve downregulation of cytokines, upregulation of MMPs, and JAK/STAT signaling mechanisms.
Division of Cardiology, Department of Pediatrics, The Children's Hospital of Philadelphia, Philadelphia, PA, USA.
Matthew J
MJ
Finley
Katherine A
KA
Clark
Ivan S
IS
Alferiev
Robert J
RJ
Levy
Stanley J
SJ
Stachelek
D000293:Adolescent; D000328:Adult; D001769:Blood / Q000378:metabolism*; D051928:CD47 Antigen / Q000737:chemistry*; D018925:Chemokines / Q000235:genetics / Q000378:metabolism; D016207:Cytokines / Q000235:genetics / Q000378:metabolism; D015536:Down-Regulation; D005260:Female; D006801:Humans; D053614:Janus Kinase 2 / Q000235:genetics / Q000378:metabolism; D008297:Male; D020781:Matrix Metalloproteinase 1 / Q000235:genetics / Q000378:metabolism; D053509:Matrix Metalloproteinase 13 / Q000235:genetics / Q000378:metabolism; D020783:Matrix Metalloproteinase 7 / Q000235:genetics / Q000378:metabolism; D008875:Middle Aged; D011143:Polyvinyl Chloride / Q000737:chemistry; D050791:STAT Transcription Factors / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D015854:Up-Regulation; D055815:Young Adult
D051928:CD47 Antigen; D018925:Chemokines; D016207:Cytokines; D050791:STAT Transcription Factors; D011143:Polyvinyl Chloride; D053614:Janus Kinase 2; C507079:MMP13 protein, human; D053509:Matrix Metalloproteinase 13; C060932:MMP7 protein, human; D020783:Matrix Metalloproteinase 7; C522360:MMP1 protein, human; D020781:Matrix Metalloproteinase 1
Biocompatibility; Cell signaling; Gene expression; Metalloproteinase