Session: 102. Iron Homeostasis and Biology: Poster III
Hematology Disease Topics & Pathways:
Research, Fundamental Science, Genetic Disorders, Hematopoiesis, Diseases, Biological Processes, Study Population, Animal model, Pathogenesis
Complete blood count analyses in the Piezo1 R2482H strain revealed that compared to wild-type (2482R/R ) littermates, mice heterozygous for the R2482H allele (2482R/H) showed significantly elevated reticulocytes (%RETIC), but showed similar hemoglobin (HGB), hematocrit (HCT), red blood cell count (RBC), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), and red cell distribution width (RDW), compatible with a compensated hemolytic defect. Mice homozygous for the R2482H allele (2482H/H) showed more severe hematological abnormalities than 2482R/H mice. Reticulocytes and RDW in 2482H/H mice were significantly higher than in 2482R/H mice, and 2482H/H mice showed mean HGB and HCT that were lower than 2482R/R controls.
Compared to either 2482R/R or 2482R/H mice, 2482H/H mice also showed significantly greater spleen weight/body weight ratio, which was accompanied by histological evidence of increased extramedullary hematopoiesis (EMH) in spleen. Total non-heme iron content of spleen (calculated as spleen non-heme iron concentration multiplied by spleen weight) was significantly higher in 2482H/H mice compared to 2482R/R controls. Compared to wild-type spleens, both 2482R/H and 2482H/H spleens showed qualitatively greater non-heme iron in red pulp macrophages by Prussian blue stain, compatible with greater red blood cell turnover. Additionally, in males, liver non-heme iron concentration was significantly higher in 2482H/H mice than in either 2482R/R or 2482R/H mice. Heart weight/body weight ratio was significantly higher in 2482H/H mice than in 2482R/R controls.
Characterization of the M2241R mouse strain revealed that compared to wild-type (2241M/M) littermates, 2241M/R heterozygotes showed significantly elevated %RETIC and maintained normal HGB, HCT, and red cell indices. Mice homozygous for the M2241R allele (2241R/R) showed significantly higher %RETIC than 2241M/R mice. However, %RETIC, spleen weight/body weight ratio, and splenic total non-heme iron content in 2241R/R mice remained significantly lower than their respective levels in mice homozygous for the R2482H allele (2482H/H). Additionally, unlike 2482H/H males, 2241R/R males did not show liver iron loading. No significant effects of the M2241R or R2482H allele were detected on serum transferrin saturation or serum hepcidin levels.
In summary, we demonstrate that different gain of function alleles of Piezo1 produce different degrees of hematological defects and iron loading in liver and spleen. Interestingly, in contrast to our results, elevated liver iron content was not detected in young adult mice homozygously expressing a conditional Piezo1 R2482H allele under the constitutive Cmv-Cre in a prior study (Ma S, Cell 2021), which might reflect differences in iron quantification methods or in the different Piezo1 alleles themselves. Future studies will be of interest to determine if the phenotypic differences observed between the R2482H and M2241R strains reflect mutation-dependent differences in PIEZO1 function in erythrocytes, macrophages, hepatocytes, and/or other cell types.
Disclosures: No relevant conflicts of interest to declare.