1. CD-RAP and Palovarotene Lab Meeting 3/18/2014

2. Palovarotene A retinoic acid receptor gamma (RARγ) agonist Inhibits cartilage formation During development, new bone formation is stimulated by a drop in retinoid signaling RARγ agonists maintain retinoid signaling preventing new bone formation Has been shown to prevent HO formation

3. B 01.472.94 Palovarotene (mg/kg) C A 01.472.94 Palovarotene (mg/kg) n = 16 n = 8 n = 16 n = 8n = 9

4. CD-RAP CD-Rap is expressed during chondrogenesis A plasma biomarker for cartilage formation – Observed to be elevated in patients with: Neurofibromatosis type I Rheumatoid Arthritis During fracture healing Marathon runners

5. Rationale – A circulating biomarker provides an additional metric for the efficacy of a treatment – A particular drug may not improve function but may result in lower biomarker levels suggesting that the class of drug may hold promise

6. Methods Plasma samples from mice and patients ELISA – Mouse: Kit from CUSABIO – Human: Kits from Roche and CUSABIO

7. D0D2D5D7D10D14 CON444448 PAL2.934458 * * Palovarotene treatment results in lower CD-RAP levels

8. Human CD-RAP ELISA

9. Conclusion and continuing work CD-RAP shows promise as a circulating biomarker for HO formation Repeat mouse CD-RAP ELISA – Increase number of samples to 8 at each time point Repeat human CD-RAP ELISA – More samples for each group – Repeat with first kit tested at higher plasma concentration