siRNA Dynamic PolyConjugates for the Targeting of Hepatocyte HAMP Genes as Potential Treatment for Anemia of Inflammation: A Research Protocol

Introduction: Anemia of inflammation (AI) is a condition caused by iron sequestration from invading pathogens, which is primarily caused by hepcidin upregulation. This results in low serum iron levels. The objective of this research protocol is to evaluate the potential of small interfering RNA (siRNA) Dynamic PolyConjugates (DPCs) in decreasing hepatic hepcidin expression for AI treatment. Methods: DPCs carrying Hepcidin Antimicrobial Peptide (HAMP) gene siRNA will be synthesized and injected into the tail veins of AI-induced mice on a standardized low-iron diet. Various experiments will then be conducted to verify that siRNA DPCs specifically target hepatocytes without causing significant toxicity. To evaluate the treatment’s efficacy, HAMP mRNA and serum iron levels will be measured using Reverse Transcription Quantitative Real- time Polymerase Chain Reaction (RT-qPCR) and a common calorimeter method, respectively. These measurements will determine the potential of siRNA to silence hepatic hepcidin expression and its resulting ability to increase serum iron levels. Results: It is anticipated that successful targeting of siRNA DPCs to hepatocytes will be confirmed through immunofluorescence and that toxicity levels induced by the treatment will be statistically insignificant. Moreover, we expect lower HAMP mRNA levels and thus higher serum iron concentrations in the experimental group compared to the control. Discussion: Hepatocyte-specific delivery of the siRNA DPC with minimal toxicity and effective silencing of the HAMP gene would deem this delivery vehicle to be a notable candidate in treating AI compared to other current conventional treatments. Certain limitations include confounding variables and potential toxicity, which should be further considered. Conclusion: Future implications of this study include human testing of siRNA DPC administration in AI patients as well as using DPCs conjugated to other siRNAs in the potential treatment of other gene-related pathologies associated with abnormal upregulation of specific proteins.