Hepcidin‐Based Nanocomposites for Enhanced Cancer Immunotherapy by Modulating Iron Export‐Mediated N6‐Methyladenosine RNA Transcript

Abstract

Despite recent advances in targeted therapies and immunotherapies in acute myeloid leukemia, patient prognosis is still dismal given the high frequency of refractory cases or post‐remission relapse. Ferrotherapy, a novel anti‐cancer treatment strategy using iron‐based compounds, can mechanistically bypass the chemoresistance of cancer cells. However, cancer cells often overexpress the transmembrane iron exporter ferroportin that expels accumulated iron into the plasma, which can potentially limit the efficacy of ferrotherapy and lead to systemic toxic effects. Here, a ferro‐therapeutic nanoplatform is designed using fusing hepcidin and leukemia cell‐membrane vesicles on gold nanoparticles (AuNPs)‐loaded hollow mesoporous Prussian Blue (AuPB@LMHep). AuPB@LMHep specifically targeted leukemia cells through ferroportin and is subsequently internalized and disintegrated into iron and AuNPs through autophagy. The accumulation of iron due to blockade of ferroportin‐mediated iron efflux and the depletion of glutathione by the AuNPs triggered ferroptosis. Furthermore, iron accumulation inactivated the endogenous iron‐dependent m6A demethylase, thereby increasing global m6A RNA modification and suppressing multiple oncogenes. Finally, AuPB@LMHep enhanced the immune response to anti‐programmed death ligand1 treatment via increasing cytotoxic tumor‐infiltrating T cells. Pre‐clinical findings provide the proof‐of‐concept of hepcidin‐based nanocomposite as an “epigenetic drug” and immunotherapeutic agent for treating leukemia.

Glutathione‐Bioimprinted Nanoparticles Targeting of N6‐methyladenosine FTO Demethylase as a Strategy against Leukemic Stem Cells

“…M 6 A Dot Blot Assay: M 6 A dot blot assay was performed as described previously. [28,55] The suitably treated cells were harvested at 1000 g for 3 min, and the pellet was homogenized with 1 mL Trizol solution. The homogenates were collected in a 1.5 mL RNA-free EP tube, and 200 µL chloroform was layered on top.…”

“…In Vivo Biocompatibility: In vivo biocompatibility was performed as described previously. [28,29] Briefly, Male Balb/c mice (4-6 weeks old, n = 3) were intravenously injected with GNPIPP12MA (200 mg mL −1 ) or PBS, and euthanized after 60 days. Approximately 500 µL blood were drawn from each mouse for routine hematological and biochemical tests.…”

Gold Nanorods Exhibit Intrinsic Therapeutic Activity via Controlling N6-Methyladenosine-Based Epitranscriptomics in Acute Myeloid Leukemia

“…Cellular uptake assay was carried out according to the previous method. 69 Briefly, cells were seeded in 6-well plates, and after 18 h of culture, the cells were treated with Cy3-labeled nanoparticles. The cells were fixed and observed under a confocal fluorescent microscope.…”

“…The fluorescence quenching experiment was carried out according to the previous method. 69 Briefly, GNRa-GSP12 was incubated with varying amounts of GSH−rhodamine. The GNR-conjugated GSH molecules were detected using a fluorescence spectra (Shimadzu RF-5301PC).…”

“…The fluorescence quenching experiment was carried out according to the previous method . Briefly, GNRa-GSP12 was incubated with varying amounts of GSH–rhodamine.…”