Bulk RNA-seq detects cMPC HO signature in hip arthroplasty patients

Heterotopic ossification (HO), the aberrant bone in soft tissues, is one of the most debilitating complications associated with severe burn and traumatic injuries as well as joint replacement surgeries due to its insidious development. Currently, no technologies exist to either support early HO detection or to guide early prophylactic strategies. Furthermore, no technologies exist to assess treatment efficacy. In this study, we used microfluidic iChip, designed to isolate circulating rare non-hematopoietic cells from whole blood, to isolate and analyze circulating mesenchymal progenitor cells (cMPCs) released in patients that underwent hip artroplasty. RNA sequencing of cMPCs revealed the unique expression of HO-associated MPC genes observed post operation day 1, weeks earlier than gold standard radiographic diagnostic strategies. Overall design: iChip enriched cells were lysed with RLT buffer (Qiagen Micro kit) and stored at -80C. Before proceeding to library preparation, RNA was purified with Qiagen micro kit. Total RNA libraries were prepared using Seqwell Rapid kit using manufacturer instructions.