P2RY14 cAMP signaling regulates Schwann cell precursor self-renewal, proliferation, and nerve tumor initiation in a mouse model of neurofibromatosis
- Division of Experimental Hematology and Cancer Biology, Cancer & Blood Diseases Institute, Cincinnati Children's Hospital Medical Center, United States
- Neuroscience Graduate Program, University of Cincinnati College of Medicine, United States
- Department of Pediatrics, University of Cincinnati College of Medicine, United States
- Molecular and Developmental Biology, Cincinnati Children’s Hospital, United States
- Hoxworth Blood Center, College of Medicine, University of Cincinnati, United States
- Department of Neurosurgery, Mayo Clinic, United States
Figures
Figure 1
P2RY14 is expressed in human neurofibromas and promotes Schwann cell precursor (SCP) self-renewal in vitro.
(A) Microarray heatmap shows P2ry14 receptor expression in p75+/EGFR+ SCP-like tumor-initiating cells derived from human plexiform neurofibroma tumor cells compared to p75+/EGFR- SCP-like cells. (B) …
Figure 2 with 3 supplements
Mouse Nf1 mutant Schwann cell precursors (SCPs) use P2RY14 signaling to regulate self-renewal.
(A) Quantification of percent of sphere forming cells in mouse wild-type (WT) and Nf1-/- SCPs treated with the selective P2ry14 inhibitor (300 nM …
Figure 2—figure supplement 1
Photomicrographs of three different experiments in which wild-type (WT) and Nf1-/- Schwann cell precursors (SCPs) were treated with 4-[4-(4-piperidinyl)phenyl]-7-[4-(trifluoromethyl)phenyl]-2-naphthalenecarboxylic acid (PPTN), shP2ry14 and rolipram.
(A) Photomicrographs of WT and Nf1-/- mouse Schwann cell (SC) spheres treated with the selective P2ry14 inhibitor (PPTN; 300 nM). (B) Photomicrographs of WT and Nf1-/- mouse SC spheres treated with …
Figure 2—figure supplement 2
4-[4-(4-Piperidinyl)phenyl]-7-[4-(trifluoromethyl)phenyl]-2-naphthalenecarboxylic acid (PPTN) dose-response analysis in wild-type (WT) and Nf1-/- Schwann cell precursors (SCPs).
(A) Photomicrographs of WT and Nf1-/- mouse Schwann cell (SC) spheres treated with the P2ry14 inhibitor (PPTN; 30 nM). (B) Photomicrographs of WT and Nf1-/- mouse SC spheres treated with the P2ry14 …
Figure 2—figure supplement 3
Photomicrographs and quantification analysis of two biological replicates of shP2ry14 in wild-type (WT) and Nf1-/- Schwann cell precursors (SCPs).
(A) Photomicrographs of WT and Nf1-/- SCP treated with shP2ry14 (64). (B) Quantification of the percent of sphere forming cells in WT and Nf1-/- SCP after treatment with shP2ry14 (64) (n = 3; …
Figure 3
P2RY14 is expressed in Schwann cell precursors (SCPs) and Schwann cells (SCs) in vivo in a mouse model of neurofibromatosis.
(A) Schematic of generation of neurofibroma mice breeding of P2ry14-/- mice with Nffl/fl mice to obtain P2ry14+/-; Nf1fl/fl;DhhCre and P2ry14-/-; Nf1fl/fl;DhhCre littermates after several crosses. (B…
Figure 4 with 1 supplement
P2RY14 deletion in a mouse model of neurofibroma increases survival and delays neurofibroma initiation.
(A) Kaplan-Meier survival plot of Nf1fl/fl;DhhCre (red line; n = 8); P2ry14+/-; Nf1fl/fl;DhhCre (black line, n = 14); P2ry14-/-; Nf1fl/fl;DhhCre (blue line; n = 13); Nf1fl/+ control (green line, n = …
Figure 4—figure supplement 1
Tumor dissection of Nf1fl/fl;DhhCre and P2ry14-/-;Nf1fl/fl;DhhCre mice at 4 months and immunostaining analysis of 4- and 7-month sciatic nerve and tumors.
(A) Representative image of gross dissection of Nf1fl/fl;DhhCre and P2ry14-/-;Nf1fl/fl;DhhCre mice at 4 months. (B) Neurofibroma tumor number quantification at 4 months of age (unpaired t-test …
Figure 5
P2RY14 deletion improves nerve ultrastructure.
(A) Electron micrograph of saphenous nerve of 4-month-old wild-type (WT), Nf1fl/fl;DhhCre, P2ry14-/-; Nf1fl/fl;DhhCre and P2ry14-/- mice. (B) Electron micrograph of saphenous nerve of 7-month-old WT,…
Figure 6
Cyclic AMP (cAMP) increase in a mouse model of neurofibromatosis either by rolipram or P2RY14 inhibitor treatment decreases Schwann cell (SC) proliferation.
(A) Rolipram drug treatment experimental design. (B) Tumor lysates of vehicle and rolipram treated Nf1fl/fl;DhhCre mice show changes in p-PKA substrate. (C) Ki67 staining at 9 months of age in …
Additional files
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Transparent reporting form
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Source data 1
Non-annotated original figures.
- https://cdn.elifesciences.org/articles/73511/elife-73511-data1-v2.zip
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Source data 2
Annotated original figures.
- https://cdn.elifesciences.org/articles/73511/elife-73511-data2-v2.zip
