Patients suffering from Sickle cell disease (SCD) present with multifactorial pathology, and a detailed understanding of it may help to develop novel therapeutics. In this study, the plasma elemental (24Mg, 44Ca, 57Fe, 63Cu, 66Zn, 77Se, 85Rb, 208Pb, and 39K) levels of SCD patients (n=10, male: 50%) and control groups (trait and healthy; n=10 each; male: 50%) were profiled using inductively coupled plasma mass spectrometry (ICP-MS). Additionally, comparative global erythrocyte metabolomics of SCD (n=5, male:100%) and healthy controls (n=5, male:100%) were carried out using liquid chromatography-mass spectrometry (LC-MS). SCD patients had higher plasma 24Mg, 44Ca, 66Zn, 208Pb, and 39K levels and lower levels of 57Fe, 77Se, and 85Rb compared to controls. These changes in elemental levels with a decreased Rb/K ratio, may explain the observed frequent hemolysis and severe dehydration with oxidative stress in the SCD group. Mass spectrometry analysis of RBCs (red blood cells) of SCD (n=5) and healthy controls (n=5) identified 442 unique metabolic features which separately clustered both the study groups in principal component analysis (PCA). A set of 136 features showed differential (p<0.05; log2fold change>+1 or -1) regulation and was involved in D-Glutamine/D-glutamate, Sphingolipid, Arginine biosynthesis, Glutathione and Glycine, serine and threonine metabolism. Interestingly, higher pyroglutamic acid levels were observed in the SS-RBCs (sickle shaped-RBC) indicating a perturbed gamma-glutamyl pathway in SCD patients. Supplementation of the depleted trace metals and targeting the perturbed metabolic pathways in the RBCs of SCD patients provides avenues for alternate therapeutics development.

The authors have declared no competing interest.

SK acknowledges the financial support from DBT, Government of India through extramural projects [BT/PR8391/BRB/10/1231/2013; BT/COE/34/SP15246/2015 and BT/PR13531/MED/30/1523/ 2015]. S Kundu also acknowledges financial support from the University of Delhi (R & D Grant; Institution of Eminence grant IOE/FRP/LS/2020/ 27); UGC, Government of India (SAP program) and DST, Government of India (PURSE Program). RN acknowledges Core support from the ICGEB New Delhi Component and project support from Department of Biotechnology New Delhi. SB acknowledges Department of Biotechnology, Government of India for Research Fellowships, ASR is thankful to the Government of Odisha for the Biju Patnaik Research Fellowship.

I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained.

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The details of the IRB/oversight body that provided approval or exemption for the research described are given below:

The IEC/IRBs of the two institutions involved in the study duly approved the protocols, procedures, and data-sharing methods for this project.They constantly monitored the protocols, subjects and recruitment procedures involved. 1. The ethics committee/IRB of Sri Sri University and Hospital, Cuttack, Odisha (SSCASRH/IEC/006/21) gave ethical approval for this work. 2. The ethics committee/IRB of University of Delhi South Campus, Delhi, India (UDSC/IEC/2021/Project/5.10.2021/4) waived ethical approval for this work. I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients, or participants themselves) outside the research group so cannot be used to identify individuals.

I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients or participants themselves) outside the research group so cannot be used to identify individuals.

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All data produced in the present study are available upon reasonable request to the authors