In the present study, we investigated the cooperative role of the TRX/GSH system during the systemic iron accumulation in mouse kidney tissue. Iron accumulation was induced in mice and the total iron content of mouse kidney tissue was determined by spectroscopic method. The expression of Fth1, Fpn1, and Hamp genes were analyzed using qPCR. The results obtained show that the iron overload model is formed. Subsequently, the impact of iron toxicity on Trx system members (Trx1, TrxR1, and Txnip) was examined at the gene and protein expression level, and enzyme activity. Iron toxicity unaffected the Trx system at the gene expression level, but activation of the Trx system was observed at the protein expression level and enzyme activity. Additionally, MDA level, TAS, TOS, and OSI were measured in mouse kidney tissue where oxidative stress was induced by iron toxicity. Finally, the effect of iron supplementation on the activities of GSH system enzymes (GPx, GR, GST) and other enzymatic antioxidants (SOD, CAT) was determined by spectroscopic method. This study demonstrated that SOD and CAT, in particular, protect mouse kidney tissue from iron overload by cooperating with the TRX system against iron toxicity. The effect of iron toxicity on kidney tissue has been shown to differ from that observed in heart and liver tissues in previous studies.

Modulation of the E3-ubiquitin ligase RNF217 alters ferroportin levels in enterocytes and iron loading in murine hemochromatosis

Liver iron overload, a contributing factor to liver damage, is frequently observed in advanced alcoholic liver disease (ALD). Genetic factors may influence the progression of ALD. This study explored the involvement of hemochromatosis-related genes in liver iron deposition among ALD patients. Clinical data from 97 ALD patients revealed that those with liver iron overload had worse liver function and prognosis. Among 43 patients who underwent exon sequencing of the HFE, HJV, HAMP, TFR2, and SLC40A1 genes, the TFR2 p.A75V mutation emerged as a potential contributor to iron overload. To further investigate, we developed a novel mouse model of ALD with liver iron overload. Administration of recombinant AAV carrying the wild-type TFR2 gene significantly reduced iron deposition in hepatocytes, whereas the TFR2 p.A75V mutation did not alleviate hepatic iron overload. This mutation impairs HAMP induction by disrupting the ERK pathway, likely due to abnormal cytoplasmic localization of the TFR2 protein. In conclusion, the TFR2 p.A75V mutation in ALD may decrease hepatocyte sensitivity to iron stimulation by inhibiting HAMP expression via the ERK pathway, thereby exacerbating iron overload both in vivo and in vitro. KEY MESSAGES: The study identifies the TFR2 p.A75V mutation as a key genetic factor worsening liver iron overload in ALD patients, linked to poorer liver function and prognosis. A novel ALD mouse model with iron overload demonstrates that the TFR2 p.A75V mutation impairs iron regulation. TFR2 p.A75V mutant disturbs the ERK pathway in HAMP induction and causes abnormal cytoplasmic localization of the TFR2 Protein.

ABSTRACTAimAtezolizumab combined with bevacizumab is the standard first‐line therapy for unresectable hepatocellular carcinoma; however, predictive biomarkers of therapeutic response remain undefined. We aimed to identify molecular features associated with therapeutic efficacy to develop personalized treatment strategies.MethodsTranscriptomic analyses were performed using public RNA‐sequencing datasets of patients with hepatocellular carcinoma receiving anti‐PD‐L1‐based therapy, comparing responders (complete response/partial response) with non‐responders (stable disease/progressive disease). Differentially expressed genes and enriched pathways were identified using integrated differential expression and pathway analysis. For validation, RNA‐sequencing was performed on institutional tumor samples (n = 6) underwent. Immunohistochemistry was performed on resected specimens (n = 9) to evaluate CD8+ tumor‐infiltrating lymphocytes and hepcidin protein expression encoded by the HAMP gene. Group comparisons for the pre‐specified immunohistochemistry endpoints were analyzed using exact Wilcoxon rank‐sum tests with multiplicity control (Holm adjustment).ResultsAnalysis of public datasets revealed distinct expression profiles in responders, enriched in immune‐related and chemokine signaling pathways. Candidate genes, including HAMP, TAT, and HRG, were upregulated in responders. In institutional samples, HAMP expression was significantly higher in preoperatively treated tumors (p = 0.001). Immunohistochemistry demonstrated greater CD8+ tumor‐infiltrating lymphocyte density (median 36.6 vs. 5.0 cells/high‐power field; exact Wilcoxon p = 0.032) and higher HAMP immunoreactive scores (median 4 vs. 0.5; p = 0.032) in responders than in non‐responders.ConclusionsUpregulation of HAMP and hepcidin protein expression, together with increased CD8+ T‐cell infiltration, was associated with a favorable response to atezolizumab–bevacizumab in patients with hepatocellular carcinoma. HAMP may serve as a component of a composite biomarker predictive of therapeutic sensitivity, warranting validation in larger, multi‐institutional cohorts.

Involvement of estrogen-related receptors in the regulatory expression of hepcidin and their relationship to hepatitis in beef cattle.

Anemia is the most common extraintestinal comorbidity of inflammatory bowel diseases (IBDs). Inflammatory cytokines are produced and exert their effects within the bowel microenvironment of IBD patients and, among them, Interleukin 6 (IL-6) induces the expression of hepcidin, encoded by the HAMP gene, contributing to the development of anemia. Enterodiol (END) and Enterolactone (ENL) are enterolignans that are known for their nutraceutical, anti-inflammatory, anti-oxidant, and estrogenic properties. Here, we used the Caco-2 cell line as an in vitro model of bowel disease to evaluate the potential nutraceutical effects of enterolignans on iron metabolism under an inflammatory stimulus induced by IL-6. As is known, IL-6 treatment induces the upregulation of HAMP gene expression. Notably, both END and ENL showed a pronounced anti-inflammatory effect, lowering HAMP mRNA levels and partially counteracting the effect induced by IL-6. The expression of iron-related genes was also studied to evaluate the effects on iron metabolism. IL-6 downregulated the expression of almost all genes studied. Notably, END and ENL mitigated the effect of IL-6, ameliorating the expression level of HAMP, FTH1, and ACO1, in addition to other END- or ENL-specific effects. The results of this study evidenced the interesting anti-inflammatory properties of enterolignans, which improved the iron homeostasis during inflammation, suggesting a possible role in the management of IBD patients with anemia.

Persistent Organic Pollutants, such as polychlorinated biphenyls (PCBs), are environmental pollutants for their resistance to degradation and adverse health effects. Despite extensive toxicological data in mammalian system, the use of alternative models such as Dictyostelium discoideum offers an opportunity to dissect evolutionarily conserved molecular mechanisms underlying pollutant-induced cellular dysfunction. In this study, we used Dictyostelium to investigate the effects of PCB 138 and PCB 153 revealing, for the first time, a direct impairment of both growth and multicellular development. PCBs exposure reduced cell proliferation and led to the formation of smaller fruiting bodies. These phenotypic effects were accompanied by altered expression of iron-regulatory genes, including upregulation of abcb7 and ferroportin, and downregulation of ferritin, consistent with intracellular iron depletion confirmed by calcein assay. We used THP-1 human cells to confirm the effect of PCBs on hamp gene, supporting the relevance of iron homeostasis as a target pathway. In Dictyostelium, iron imbalance was associated with increased ROS levels, downregulation of superoxide dismutase genes, and altered mitochondrial morphology. Under starvation, PCBs-treated cells also showed transcriptional upregulation of key development genes involved in cAMP signaling (acaA, carA, regA, gtaC), while proteomic analysis revealed changes in proteins linked to cell adhesion, stress response, and development. Together these findings support a model in which PCBs induce iron efflux, oxidative stress, and disruption of developmental signaling, ultimately both proliferation and morphogenesis. This study highlights the potential of Dictyostelium discoideum as a sensitive and cost-effective model to uncover conserved cellular responses to environmental pollutants.

Malnutrition of protein and essential nutrients in children can lead to serious health problems. It significantly alters hepatic physiology and leads to impaired liver function. The present study investigated the underlying mechanism of malnutrition-induced steatohepatitis in a rat model. Weanling rats were divided into two groups. The control rats received a standard protein diet, while the other group was fed a low protein diet (LPD) for eight weeks. LPD significantly reduced the body and liver weights and altered the blood parameters. LPD resulted in elevated serum liver injury markers and lowered glucose, albumin, and total protein levels. The reduced levels of TIBC and TSI and upregulated expression of Hamp gene were observed in the LPD group. Histopathology revealed the severe fat accumulation in the hepatocytes, leading to inflammation and fibrognesis. LPD upregulated the de novo lipogenesis (Srebp1c, Fas, Acc, and Scd1) markers and oxidative stress in the hepatic tissue. The downregulation of Pgc1α, Tim23, and Tfam indicated mitochondrial dysfunction in the LPD group. Transcriptomic analysis revealed the upregulation of 7,545 genes in the LPD group mainly associated with metabolic dysfunction-associated steatotic liver disease (MASLD), beta-oxidation, AMPK signalling and oxidative phosphorylation. Hepatic lipidome revealed the elevated levels of various lipid species in the LPD group. Further, LPD altered the gut microbiome of rats and reduced the relative abundance of beneficial bacteria. The present study revealed that malnutrition induces hepatic steatoheptitis by altering the hepatic lipid metabolism and disrupting mitochondrial function and gut-liver axis.

Background: β-thalassemia major, a genetic disorder causing chronic anemia and iron overload, often leads to dysregulated hepcidin, a key iron regulator. Bacterial infections, common in these patients, may alter hepcidin expression. Object: This study aims to assess the impact of bacterial infections on serum hepcidin levels and related HAMP (rs10421768 C-582A > G) in patients with β-thalassemia major. Method: This was a cross-sectional study with prospective data collection from August to December 2024. The study included two groups: cases, consisting of patients with β-thalassemia major at the Erbil Thalassemia Center, and controls, consisting of healthy individuals unrelated to the case group. Hepcidin levels were determined using a sandwich ELISA (enzyme-linked immunosorbent assay). Bacterial identification was performed on a urine sample using the VITEK 2 system. Additionally, a genetic test was conducted using ARMS-PCR. Data were analyzed using GraphPad Prism 9.0 with t-tests for group comparison and chi-square tests for genotype distribution. Result: In 100 β-thalassemia major patients, mean hepcidin levels were higher in those with bacterial infection (62.15 ± 6.676 ng/mL) than in non-infected patients (54.65 ± 5.286 ng/mL). Gram-negative bacteria (52.55%) were more common, with Escherichia coli being most prevalent (31.5%). The HAMP gene polymorphism (rs10421768 C-582A > G) was most frequently observed as the GA genotype, which showed differences in serum hepcidin levels between healthy and thalassemia patients. Conclusion: Hepcidin levels in β-thalassemia major patients are influenced by both bacterial infections, especially Gram-negative bacteria, and HAMP (rs10421768 C.-582A>G) gene polymorphism, underscoring the combined role of genetic and infection factors in iron regulation

Relevance. Iron deficiency anemia is a disease present all over the world and maintaining a high incidence rate of both general and primary morbidity despite various options for correction and prevention of this pathology. Molecular genotyping opens up new possibilities for understanding the pathogenetic mechanisms of disease development, as well as for finding a more targeted and personalized approach to therapy. The aim of the study was to determine genetic markers of the risk of iron deficiency anemia by assessing polymorphic variants of rs235756 of the BMP2 gene, rs855791 of the TMPRSS6 gene, rs104894696 of the HAMP gene, rs4820268 of the TMPRSS6 gene and rs3817672 of the TFR1 gene. Material and methods. The study included 97 patients with mild iron deficiency anemia and 120 people in the control group. Genotyping was performed by real-time polymerase chain reaction using primers and probes selected using the open online resource PrimerBlast (https://www.ncbi.nlm.nih.gov/tools/primerblast/). Results. Genetic markers of the risk of developing IDA were identified: rs235756 of the BMP2 gene, rs855791 of the TMPRSS6 gene and resistant course of IDA: rs4820268 of the TMPRSS6 gene. This allows us to recommend determining these genetic markers when examining patients with iron deficiency anemia who do not respond to therapy with iron-containing drugs. Conclusion. The identified marker profiles of IDA genotypes can optimize the approach to diagnosis, treatment and prevention of this pathology, as well as expand the range of criteria for predicting the course of the disease

Hemochromatosis is an autosomal recessive iron overload disorder. It occurs due to a failure in the hepcidin response, leading to systemic iron overload. The high iron levels in the plasma stored in various organs cause injury and permanent damage. There are two types of hemochromatosis: primary and secondary. In non- HFE hemochromatosis, mutations in the HJV, HAMP, TRF2, and SLC40A1 genes are implicated, with the associated condition classified as type I hemochromatosis. In contrast, juvenile hemochromatosis (type II hemochromatosis/ HFE II) is linked to mutations in the hemojuvelin gene or the antimicrobial peptide hepcidin. In this study, relevant literature in databases, including PubMed, MEDLINE records, Cochrane Central Register of Controlled Trials (CENTRAL), Google Scholar, and Embase, was searched. Our study inclusion criteria encompassed both experimental and observational studies or a combination of both, with data derived from the human population. The exclusion criteria included animal models, observational studies, and unpublished data. Hepcidin is usually up-regulated in response to high serum iron, but it is unexpectedly low in patients with hemochromatosis because of mutations in HFE, hemojuvelin (JH), and transferrin receptor 2 (TfR2). TfR2, expressed by hepatocytes, is mutated in hemochromatosis type III. Future research directions include exploring the molecular mechanisms underlying the effects of the TFR2 gene variant on iron homeostasis and liver damage and investigating potential therapeutic targets for treating hemochromatosis-related liver disease. Additionally, further epidemiological and modern genetic engineering studies are needed to better understand the prevalence and impact of hemochromatosis on liver health in different populations.

BackgroundAn oligopeptide hepcidin is encoded by the human HAMP gene (Hamp in mice). Its deficiency can result in iron overload, while excess may lead to iron deficiency. Hepcidin knockout mice exhibited iron accumulation in multiple tissues, accompanied by degeneration of bone microarchitecture and reduced biomechanical properties. Astronauts who are exposed to weightlessness during prolonged spaceflight experience bone loss. After space missions, an interrelation exists between iron stores and bone mineral density (BMD). Bone loss in mice due to unloading is linked to iron excess and involves hepcidin. The potential role of hepcidin in unloading-induced bone loss remains unclear.MethodsOur study conducted relevant experiments using hepcidin knockout mice and their primary osteoblasts as the research subjects. We used the hindlimb unloading (HLU) model and the random positioning machine (RPM) system to simulate weightlessness in vivo and in vitro.ResultsHLU mice exhibited reduced hepcidin levels in the serum and liver. Hepcidin knockout further diminished BMD and bone mineral content (BMC) in the femurs of HLU mice. Similarly, the bone volume fraction (BV/TV) and connectivity density (Conn.Dn) followed this downward trend, whereas trabecular separation (Tb.Sp) showed an inverse pattern. Moreover, hepcidin knockout decreased the ultimate load and elastic modulus in the tibias of HLU mice. Hepcidin knockout decreased PINP levels in the serum, a commonly used marker for bone formation, alongside elevated iron levels in the serum, liver, and bone of HLU mice. We also found higher serum MDA and SOD levels in these mice. In vitro, experimental data indicated that hepcidin knockout suppresses the osteoblastic differentiation capacity under RPM conditions. Additionally, this condition upregulates SOST protein levels and downregulates LRP6 and β-catenin protein levels in osteoblasts.ConclusionHepcidin knockout exacerbates bone loss in HLU mice, most likely due to reduced osteoblastic activity.

ObjectiveMechanism underlying the etiology of polycystic ovary syndrome (PCOS) is still debatable. Present study explores the link between iron-mediated ferroptosis and PCOS.MethodologyBlood samples were collected from 150 PCOS females along with healthy controls. Expression analysis of FTH1, NCOA4, GPX4, HAMP, A2M and HP genes was estimated by RT-qPCR. Serum was used for estimation of lipid peroxidation, peroxidase enzyme, ferritin and total protein.ResultsRelative expression of FTH1 (P < 0.05), HAMP (P < 0.01), GPX4, A2M, HP (P < 0.001) was downregulated and NCOA4 (P < 0.001) was upregulated in PCOS group compared to control. A significant difference was observed in mRNA expression of selected genes when ≤ 30year age group PCOS was compared to > 30year age PCOS group and their respective controls. Deregulation of gene expression was prominent in PCOS group with obese and overweight BMI compared to underweight and normal BMI group. Menstrual cycle length and marital status of PCOS females had no significant association with selected gene expression. Expression deregulation in targeted genes was observed in PCOS patients with complaints of either diabetes, high blood pressure or both. Increased level of lipid peroxidation, serum ferritin and total protein, while decreased peroxidase activity was observed in PCOS group (P < 0.001) compared to control.ConclusionThe present study postulated the role of iron overload in trigger of ferroptosis following elevated lipid peroxidation and low peroxidase activity. Moreover, unveil the association of genes related to iron-regulating metabolism with etiology of underlying PCOS mechanism.Graphical

Menopause is a natural biological aging process characterized by the loss of ovarian follicular function and decrease estrogen levels. These hormonal fluctuations are associated with increased iron levels, which ultimately lead to iron accumulation. This study aims to investigate the effects of Deferasirox on iron homeostasis and hematopoiesis in ovariectomized rats with iron accumulation. Sixty-four female Wistar rats were divided into eight groups and underwent ovariectomy surgery to simulate menopause. Iron accumulation was induced through the injection of ammonium ferric citrate. Deferasirox was administered at doses of 50 mg/kg and 100 mg/kg. Hematological parameters, iron profile, antioxidant markers, oxidative stress indicators, histopathological evaluation of uterine, bone, bone marrow, liver, and spleen tissues, flow cytometric analysis of hematopoietic CD markers, and relative expression of Hamp, Pu.1, Gata1, and Gdf11 genes were analyzed. Deferasirox treatment improved histopathological changes in the uterine tissue of ovariectomized rats with iron accumulation, increased the number of white blood cells, and reduced serum iron levels, TIBC, ferritin, and transferrin saturation percentage. It also increased serum antioxidant capacity and reduced oxidative stress markers. Deferasirox had a positive effect on femur bone, hematopoietic cell count, volume of hematopoietic and adipose tissues in bone marrow, extramedullary hematopoiesis in the liver and spleen, and influenced the relative expression of Hamp, Pu.1, Gata1, and Gdf11 genes related to hematopoiesis and iron metabolism. In conclusion, Deferasirox effectively manages iron homeostasis and hematopoiesis in ovariectomized rats with iron accumulation and suppresses oxidative stress.

The incidence of juvenile Hereditary Hemochromatosis caused by HAMP gene mutation is low, which is rarely reported in China. This patient took abnormal liver function as the first symptom, and was finally diagnosed by genetic testing and hepatic histopathology, and treated by venous bloodletting.

PIEZO1 encodes a mechanoreceptor, a cation channel activated by mechanical stimuli. Gain‐of‐function (GoF) variants in PIEZO1 cause dehydrated hereditary stomatocytosis (DHS), or xerocytosis, a pleiotropic syndrome characterized by anemia and iron overload. DHS patients develop hepatic iron overload independent of the degree of anemia and transfusion regimen. PIEZO1‐GoF variants suppress hepcidin expression in both hepatic cellular model and constitutive/macrophage‐specific Piezo1‐GoF mice model. Therefore, PIEZO1‐GoF variants regulate hepcidin expression by a crosstalk between hepatocytes (HCs) and macrophages with a still unknown mechanism. Transcriptomic and proteomics analysis in the human hepatic Hep3B cells engineered for the PIEZO1‐R2456H variant (PIEZO1‐KI) revealed alterations in the actin cytoskeleton regulation, MAPK cascade, and RAS signaling. These changes mainly occur through a novel key regulator, RRAS, whose protein and mRNA levels are regulated by PIEZO1 activation and inhibition. This regulation was further confirmed in C57BL/6 mouse primary HCs treated with Yoda‐1 and/or GsMTx‐4. Indeed, PIEZO1‐KI cells exhibited hyper‐activated RAS‐GTPase activity that is rescued by PIEZO1 inhibition, restoring expression of the hepcidin gene HAMP. A negative correlation between RAS signaling and HAMP regulation was confirmed by inhibiting RAS‐GTPase and MEK1‐2 activity. Conversely, rescued HAMP gene expression requires downregulation of RRAS, confirming negative feedback between RAS‐MAPK and BMP/SMADs pathways in HAMP regulation. We demonstrated that PIEZO1‐GoF variants influence the actin cytoskeleton organization by activating the hepatic RAS signaling system. Understanding the role of RAS signaling in regulating iron metabolism could pave the way for new therapeutic strategies in DHS and other conditions characterized by iron overload.

Hetrombopag, an Emerging Iron-Chelating Agent, Alleviates Systemic Iron Overload

BackgroundRheumatoid arthritis (RA) is a chronic systemic autoimmune inflammatory disorder characterized by synovial inflammation that leads to joint damage, bony erosions, and related deformities. Between 30 and 70% of RA patients will experience anemia. Early detection of anemia is of great importance. This study aimed to evaluate the serum level of hepcidin (HEP) in RA patients and to assess its relation to disease activity and anemia. The current cross-sectional study included 44 cases with RA in addition to 44 healthy controls. The disease activity in the RA patient was assessed by using the disease activity score (DAS) 28 score-CRP. The serum levels of HEP and ferritin were assessed in both groups using enzyme-linked immunosorbent assay (ELISA) technique.ResultsHepcidin level in the RA group was statistically significantly higher as compared to the control group (p = 0.001). The prevalence of Anemia of chronic disease (ACD) was 40.9%, and iron deficiency anemia (IDA) was 27.3% which accounted for 68.2% of the total anemia cases. The HEP level was statistically significantly higher in the RA patients with ACD than those without anemia (P = 0.028), RA patients with IDA (P < 0.001), and control group (P < 0.001). There was a statistically significant positive correlation between HEP level and serum ferritin level (p = 0.005). HEP level was significantly and inversely correlated with hemoglobin (Hb) in patients with ACD. Serum HEP level is higher in RA patients with high disease activity than those with moderate activity, low activity, and patients in remission (p = 0.380). However, the difference was not statistically significant. The best cutoff point of HEP level to identify RA patients from healthy controls was > 355.5 Pg/ml. This point showed moderate sensitivity (70.5%) with moderate specificity (63.6%) with a statistically significant value.ConclusionsWe found the anemia, and particularly ACD, is more common in RA patients. In RA patients with ACD, serum HEP levels were considerably higher. Although serum HEP showed no diagnostic significance when it came to evaluating disease activity, it could be a dependable non-invasive biomarker for the diagnosis of various forms of anemia in RA patients.

Exercise-induced inflammation can influence iron metabolism. Conversely, the effects of vitamin D3, which possesses anti-inflammatory properties, on ultramarathon-induced heart damage and changes in iron metabolism have not been investigated. Thirty-five healthy long-distance semi-amateur runners were divided into two groups: one group received 150,000 IU of vitamin D3 24 h prior to a race (n = 16), while the other group received a placebo (n = 19). Serum iron, hepcidin (HPC), ferritin (FER), erythroferrone (ERFE), erythropoietin (EPO), neopterin (NPT), and cardiac troponin T (cTnT) levels were assessed. A considerable effect of ultramarathon running on all examined biochemical markers was observed, with a significant rise in serum levels of ERFE, EPO, HPC, NPT, and cTnT detected immediately post-race, irrespective of the group factor. Vitamin D3 supplementation showed a notable interaction with the UM, specifically in EPO and cTnT, with no other additional changes in the other analysed markers. In addition to the correlation between baseline FER and post-run ERFE, HPC was modified by vitamin D. The ultramarathon significantly influenced the EPO/ERFE/HPC axis; however, a single substantial dose of vitamin D3 had an effect only on EPO, which was associated with the lower heart damage marker cTnT after the run.

Iron-refractory iron deficiency anaemia (IRIDA) is a rare autosomal recessive disorder, distinguished by hypochromic microcytic anaemia, low transferrin levels and inappropriately elevated hepcidin (HEPC) levels. It is caused by mutations in TMPRSS6 gene. Systematic screening of 500 pregnant women with iron deficiency anaemia having moderate to severe microcytosis with no other causes of anaemia were enrolled to rule out oral iron refractoriness. It identified a final cohort of 10 (2.15% prevalence) individuals with IRIDA phenotype. Haematological and biochemical analysis revealed significant differences between iron responders and iron non-responders, with iron non-responders showing lower haemoglobin, red blood cell count, serum iron and serum ferritin levels, along with elevated HEPC (9.47 ± 2.75 ng/mL, p = 0.0009) and erythropoietin (4.58 ± 4.07 µ/mL, p = 0.0196) levels. Genetic sequencing of the TMPRSS6 gene in this final cohort identified 10 novel variants, including seven missense and three frame-shift mutations, with four missense variants showing high functional impact defining the IRIDA phenotype. Structural analysis revealed significant damage caused by two variants (p.L83R and p.S235R). This study provides valuable insights into IRIDA among pregnant women in the Indian subcontinent, unveiling its underlying causes of unresponsiveness, genetic mechanisms and prevalence. Furthermore, research collaboration is essential to validate these findings and develop effective treatments.