Dietary ω-3 Fatty Acid Supplementation Improves Murine Sickle Cell Bone Disease and Reprograms Adipogenesis

Abstract

Sickle cell disease (SCD) is a genetic disorder of hemoglobin, leading to chronic hemolytic anemia and multiple organ damage. Among chronic organ complications, sickle cell bone disease (SBD) has a very high prevalence, resulting in long-term disability, chronic pain and fractures. Here, we evaluated the effects of ω-3 (fish oil-based, FD)-enriched diet vs. ω-6 (soybean oil-based, SD)- supplementation on murine SBD. We exposed SCD mice to recurrent hypoxia/reoxygenation (rec H/R), a consolidated model for SBD. In rec H/R SS mice, FD improves osteoblastogenesis/osteogenic activity by downregulating osteoclast activity via miR205 down-modulation and reduces both systemic and local inflammation. We also evaluated adipogenesis in both AA and SS mice fed with either SD or FD and exposed to rec H/R. FD reduced and reprogramed adipogenesis from white to brown adipocyte tissue (BAT) in bone compartments. This was supported by increased expression of uncoupling protein 1(UCP1), a BAT marker, and up-regulation of miR455, which promotes browning of white adipose tissue. Our findings provide new insights on the mechanism of action of ω-3 fatty acid supplementation on the pathogenesis of SBD and strengthen the rationale for ω-3 fatty acid dietary supplementation in SCD as a complementary therapeutic intervention.

Transfusional Approach in Multi-Ethnic Sickle Cell Patients: Real-World Practice Data From a Multicenter Survey in Italy

“…Of note, we found a subset of patients still symptomatic for acute SCD-related clinical manifestations under treatment with HC, highlighting the biocomplexity of SCD that lies beneath red cell sickling and dehydration ( 38 – 44 ). Indeed, HC partially acts on sickle cell-related inflammatory vasculopathy and chronic inflammation ( 4 , 45 – 47 ).…”

Expression of FBXW11 in normal and disease‐associated osteogenic cells

“…Proteins were separated by using SDS‐PAGE. Western blot analyses were performed as previously reported 27 . Briefly, proteins were extracted with the RIPA buffer (Thermo Fisher Scientific).…”

“…Then, the proteins were transferred onto polyvinylidene difluoride membranes (Thermo Fisher Scientific) and probed with primary antibodies (RUNX2; Cell Signalling; 8486) (FBXW11; PA5‐109715; Invitrogen) (BetaCatenin; PA5‐19469; Invitrogen) (β‐actin; BA3R) (Thermo Scientific) and secondary antibodies anti‐mouse (Cell Signalling; 7076) and anti‐rabbit (Cell Signalling; 7074). Signals were detected with a method through the chemiluminescence reagent (ECL; Millipore), as previously reported 27 . LAS4000 Digital Image Scanning System (GE Healthcare) was used to acquire the images and densitometric analyses were performed by ImageQuant software (GE Healthcare) as previously reported 27 …”

“…Western blot analyses were performed as previously reported. 27 Briefly, proteins were extracted with the RIPA buffer (Thermo Fisher Scientific). BCA assay (Thermo Fisher Scientific) was used to quantify the proteins.…”

“…Signals were detected with a method through the chemiluminescence reagent (ECL; Millipore), as previously reported. 27 LAS4000 Digital Image Scanning System (GE Healthcare) was used to acquire the images and densitometric analyses were performed by ImageQuant software (GE Healthcare) as previously reported. 27 …”