Department of Genetic Engineering and Biotechnology

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Remedial Effects of Homeopathic Drugs against Chronic Suppuration of Ear Caused by Pathogenic Microorganisms
(2019-10-07) Hasan, Nazmul
Chronic Suppurative Otitis Media (CSOM) is known as one of the most common infectious disease and is characterized by a persistent discharge from the middle ear through a perforated tympanic membrane. CSOM is considered as one of the most common cause of hearing impairment in the developing countries. Use and misuse of antibiotics for treating CSOM promotes antibiotic resistance leading to accelerated public health risks. Homeopathic medicines are mostly free from side effects, and therefore, this study was performed to evaluate the effects of homeopathic medicines for remedies of CSOM symptoms. A total of 70 subjects were included in this study; among them, 40 were suffering from CSOM and 30 were healthy control. The CSOM patients were randomly assigned for daily administration of homeopathic medicines such as tellurium metallicum in 6C potency (n=17), mercurius solubilis in 200C potency (n=13 ) and calcarea sulphurica in 30C potency (n=10 )for 8 weeks. Aural swab of the patients were examined to assess microbiological infection. Identification of the isolated microorganisms such as Staphylococcus sp., E coli, klebsiella sp. was confirmed with Analytical Profile Index (API ). The homeopathic medicines did not show significant antibacterial activity when assessed by agar disc diffusion; however, they reduced average microbial load significantly. Ear suppuration was prevented in all of the patients treated with the medicines. The patients were also examined with otoscope to observe the level of their tympanic membrane perforation and the healing of the perforated membrane after treatment. Among the seventeen (17) patients suffering from CSOM associated with membrane perforation, 9 patients’ (52.9% ) membranes were healed when treated with tellurium metallicum. Calcarea sulphurica also healed perforated tympanic membranes of 50% patients, however, mercuris solibulis failed to trigger membrane healing in treated patients. It was next examined whether the patients also had hearing impairment compared to healthy control subjects. The average hearing thresholds of the control subjects (n=30) at 1, 4, 8 and 12 kHz frequencies were 13.55 ± 1.57, 15.58 ± 1.12, 27.58 ± 2.00 and 32.57 ± 2.45 dB SPL, respectively. When the average hearing thresholds of the CSOM patients (n=40) were measured at all of the above frequencies, the values became 30.25 ± 1.75, 36.80 ± 3.25, 50.75 ± 2.35, and 59.50 ± 2.96 dB SPL, respectively. This result indicated that the CSOM patients were suffering from hearing loss as the average hearing thresholds of the CSOM patients at all the frequencies tested were significantly higher (p<0.05)than that of the control subjects. To our surprise, treatment for 8 weeks with all three homeopathic medicines significantly reduced (p<0.05) hearing thresholds of the patients at all the frequencies. This result clearly indicated that the patients experienced dramatic improvement in hearing by the homeopathic medicines. This study, therefore, provides conclusive evidence that homeopathic medicines prevent ear suppuration in CSOM patients as well as reduce microbial load, heal perforated membrane and improve hearing.
Detection of mutation and polymorphism in marker genes of cervical carcinoma in Bangladeshi patients
(University of Dhaka, 2019-10-13) Zohura, Fatima Tuj
Cervical cancer is a gynaecologic cancer type that develops in the cervix. Annually 528,000 new cases of cervical cancer are reported with 266,000 deaths, which account for 8% mortality of all female cancer patients. Infection with specific human papillomavirus (HPV) types is considered the most severe risk factor for cervical cancer. In the context of our socioeconomic conditions, an increasing burden of this disease and high mortality rate prevail in Bangladesh. Although several researches related to the epidemiology, HPV vaccination, and treatment modalities have been carried out in our country, any research on mutation locations and frequencies in cervical cancer in Bangladesh is yet to be done. Among the high number of genes involved in different signal transduction and cell growth regulation pathways, five different genomic regions within the top three most frequently mutated genes in COSMIC database with a key role in the development of cervical cancers were selected to study mutation frequency in our patients. These genes are EGFR (Epidermal Growth Factor Receptor), KRAS (Kirsten rat sarcoma), and PIK3CA (phosphatidylinositol-4,5bisphosphate 3-kinase, catalytic subunit alpha). DNA from 46 cervical tissue samples were extracted and amplified by PCR, using 1 set of primers designed for EGFR and 2 sets of primers designed for two different regions of both PIK3CA and KRAS gene. PCR products were purified and sequenced through Sanger cycle sequencing strategy. In silico analysis was done in two steps: nucleotide sequence analysis and its corresponding amino acid analysis. The mutated protein structures were determined by homology modelling using SWISS-MODEL. In total, 39 mutations were found in 28 patient samples. Eleven different mutations (23.91%) were found in amplified EGFR gene fragments, among which 1 was common in seven patient samples. On the other hand, twenty four different mutations (52.17%) were found in PIK3CA gene fragment amplicons, among which 2 were found in more than 1 patient. Four mutations (8.7%) were found in KRAS gene fragment amplified products. Our study shows that except for KRAS, the frequency of observed mutations in our patients is higher than those reported earlier in other parts of the world. The study can be used as a basis to build a mutation database for cervical cancer in Bangladesh. With the possibility of targetable oncogenic mutations, further exploration can be oriented towards establishing future diagnostics, personalized medicine decisions, and other pharmaceutical applications for specific cancer subtypes.
Genetic diversity among Bangali and seven ethnic groups of Bangladesh based on Y-chromosome
(University of Dhaka, 2021-08-24) Hasan, Md. Mahamud
A comprehensive population genetic study was conducted using 17 Y-chromosome specific microsatellites to decipher genetic diversity of the Bangladeshi populations from a Y-chromosome perspective. Seven ethnic populations living in different parts of Bangladesh along with the mainstream Bangali population was included in this study. A total of 1,594 haplotypes were typed in 1,631 male subjects belonging to Bangali, Chakma, Tripura, Rakhine, Marma, Hajong, Manipuri, and Khasia with corresponding discrimination capacity of 97.301, 73.885, 65.563, 75.862, 77.536, 80.909, 74.809, and 81.250%, respectively. Overall, 1,359 different haplotypes were detected, of which 1,222 (89.919%) were each found in a single male-only. Observed haplotype diversity values were in a range between 0.982 (Manipuri) and 0.998 (Bangali). Bangalis have the highest average gene diversity value of 0.662 while tribal populations showed less. The pair-wise genetic distances Rst values between the studied populations and 32 worldwide reference populations revealed that the strong genetic relationship of Bangali with Nepalese (Kathmandu, Nepal), Irani (Central Iran, Iran), Iraqi (Iraq), Tamil (Tamil Nadu, India), and Bhil (Gujarat, India) populations. In addition, Chakma, Tripura, Rakhine, Marma, and Hajong were the most similar population to Tripuri (Tripura, India), Garo (Tangail, Bangladesh), Han (Beijing, China), Korean (Seoul, South Korea), Asian American (United States), and Japanese (Tokyo, Japan) populations, respectively. In contrast, Manipuri showed close genetic affinity with Tamil (Tamil Nadu, India) and Bhil (Gujarat, India) population. Moreover, Khasia lies closer to a clade consisting of Oraon (Chhattisgarh, India) and Santal (Dinajpur, Bangladesh). The studied samples analyzed were categorized into twenty different haplogroups. The Bangali showed greater frequency for haplogroups R1a, H, and L, whereas the studied seven ethnic populations exhibited higher frequency for haplogroups L, Q, R1a, and T. The Bangali contains paternal lineages from India, Pakistan, Afghanistan, and Iran of West Asia, whereas the ethnic populations contain paternal lineages from the northern and southern areas of East Asia. The study has also constructed haplotype and allele frequency data for 17 forensic identity markers and characterized the levels of populations sub-structure as well as genetic relationship within and between Bangladeshi populations.
Genetic Spectrum and Metabolic Profiling of Congenital Hypothyroid Children in Hospital Care Settings in Bangladesh
(University of Dhaka, 2021-08-26) Begum, Mst. Noorjahan
Congenital Hypothyroidism (CH) is the deficiency of thyroid hormones at birth and a major cause of mental retardation if left untreated. The frequency of CH is much higher in Bangladesh than that of the global incidences and late-diagnosis is a common scenario. There is limited data on genetic etiology of Congenital Hypothyroidism in Bangladesh and patients diagnosed at late-stage suffer from clinical complications even under treatment with levothyroxine drug. The present study aimed to investigate the genetic spectrum and metabolic profiling of Congenital Hypothyroid children of Bangladesh in order to correlate the clinical complications of CH with the blood acylcarnitines and amino acids levels. A total of 65 confirmed cases of Congenital Hypothyroid children under follow-up treatment were enrolled in this study. Among them 44 participants with thyroid Dyshormonogenesis were considered for TPO gene analysis; and 21 cases with thyroid Dysgenesis were studied for TSHR and PAX8 gene. Moreover, 107 healthy subjects were enrolled for metabolic profiling. Molecular analysis was performed using PCR followed by Sanger sequencing of TPO, TSHR and PAX8 genes. The effect of mutations on the 3D structure of the TPO monomer and dimer structures along with TSHR proteins were analyzed by Molecular Docking and Molecular Dynamics Simulations using Autodock vina/PyRx protocol and YASARA computational software. Finally, High Resolution Melt (HRM) curve analysis method was set up to detect the common mutations found in the TPO gene. For metabolic profiling, blood acylcarnitines and amino acids levels of patients were compared with healthy controls using LC-MS/MS technique. Analysis of TPO gene identified 12 mutations. In almost all patients, 4 mutations belonging to Exon 8 and Exon 12 (hotspot region) were commonly detected. Analysis of the effect of the three non-synonymous mutations c.1117G>T (p.Ala373Ser); c.1193G>C (p.Ser398Thr); and c.2173A>C (p.Thr725Pro) on the 3D structure of TPO protein revealed alterations in the binding affinities of the ligands (Heme, H2O2 and I-) for the wild type cases compared to mutant cases. A HRM (High Resolution Melt) curve analysis method was established to detect these 3 mutations in homozygous, heterozygous and wild type states. Analysis of TSHR gene on the other hand, found a mutation c.1523C>T (p.Ser508Leu) in one patient and 20 patients had c.2181G>C (p.Asp727Glu) mutation in Exon 10 of the gene. The effect of these two mutations on 3D structure of small molecules drug (MS437 and MS438) binding site was investigated and we found that the binding affinities of the small molecules were decreased in case of mutant TSHR protein compared to wild type TSHR protein. No mutation was detected in PAX8 gene. Profiling of acylcarnitines showed that the mean concentration of long chain acylcarnitines was significantly lower in the patients group than that of the Healthy Controls (HCs) group (with a P=0.0014). Also, the indicator ratio of β-oxidation rate, namely C0/(C16+C18) was significantly higher in the patient group than that of the HC group (P<0.0001), suggesting a low activity of CPT-I enzyme in the patients. Furthermore, the mean plasma TG level of the patients was significantly higher than that of the HC group (P=0.02) suggesting that the patients might have been suffering from fatty liver. Moreover, in terms of the analyzed amino acids profile, 9 out of 15 amino acids were significantly decreased in the patients compared to the healthy controls (P<0.0008). Serum albumin levels were also significantly (P<0.0001) lower in the patients group compared to the HCs, suggesting that there might be a decreased rate in overall protein metabolism. This study is the first of its kind that demonstrated the genetic basis of Congenital Hypothyroidism in Bangladeshi patients. The correlation between mutation and functional activity of specific proteins were also investigated using several in silico tools. In conclusion, metabolic profiling combined with mutation analysis might help to better understand the disease severity and to modify the treatment strategy of congenital hypothyroid children in Bangladesh.
Inborn errors of metabolism screening and profiling of amino acids, acylcarnitines and glucose-6-phosphate dehydrogenase deficiency in Bangladesh population
(University of Dhaka, 2021-08-29) Sarker, Suprovath Kumar
Similar to other lower and lower middle income countries, Bangladesh is facing the accelerated demographic shift from communicable diseases to non-communicable diseases (NCDs) including genetic disorders (GDs). Approximately 10,000 single-gene disorders have been identified which affects millions of people worldwide. One of the major categories of phenotypically and genotypically heterogeneous group of genetic disorders are inborn errors of metabolism (IEM). Liquid Chromatography tandem mass spectrometry (LC-MS/MS) is commonly used for the diagnosis of more than 30 IEM. Accurate and reliable diagnosis of IEM by quantifying amino acids (AAs) and acylcarnitines (ACs) using LC-MS/MS systems depend on the establishment of age-specific cut-off values of the analytes for the population under investigation. Thus, the first objective of the study was to (1) determine the age-specific cut-off values of AAs and ACs in Bangladesh and (2) validate the LC-MS/MS method for diagnosis of the patients with IEM. A total of 570 healthy participants were enrolled in this study. They were divided into 3 age groups, namely, (1) newborns (1-7 days), (2) 8 days–7 years, and (3) 8–17 years, to establish the age-specific cut-offs for AAs and ACs. In addition, 273 suspected patients with IEM were enrolled to evaluate the reliability of the established cut-off values. Among these patients, nine cases came out as screening positive by LC-MS/MS and seven cases were confirmed by urinary GC-MS analysis including 3 cases with phenylketonuria, 1 with methylmalonic acidemia, 1 with isovaleric acidemia, 1 with citrullinemia type II, and 1 with carnitine uptake defect. Two borderline positive cases with medium-chain acyl-CoA dehydrogenase deficiency were negative by urinary GC-MS analysis. The second objective of the study was to detect mutations in glucose-6-phosphate dehydrogenase (G6PD) gene that are responsible for G6PD deficiency in Bangladeshi individuals because G6PD deficiency is also very common like IEM in the Indian subcontinent. Among 121 clinically suspected patients, 12 (11 males and one female) patients were found to be G6PD deficient, suggesting the frequency of G6PD deficiency among suspected patients is 9.9%. Sanger sequencing revealed c.C131G substitution (exon-3: Ala44Gly) in six samples, c.G487A substitution (exon 6: Gly163Ser) in five samples and c.G949A substitution (exon-9: Glu317Lys) in the coding sequence in one sample. From the study, it appears that Ala44Gly and Gly163Ser are the most common G6PD mutations in Bangladesh The third objective of the study was to establish a real-time PCR-based approach called high-resolution melt (HRM) analysis as a rapid and reliable method for the detection of G6PD variants in heterozygous females. Although hemizygous males and homozygous females are easily detected by the conventional G6PD enzyme assay method, the heterozygous state can be missed by the conventional methods as the mosaic population of both normal and deficient RBCs circulate in the blood. Sixty-three clinically suspected females were evaluated for G6PD status using both enzyme assay and HRM analysis. Four out of sixty-three participants came out as G6PD deficient by the enzyme assay method, whereas the HRM approach could identify nine participants with different G6PD variants, namely one homozygous and eight heterozygous. In conclusion, along with the establishment of a validated LC-MS/MS method for the quantitation of amino acids and acylcarnitines from the DBS cards, the study also demonstrates the presence of predominant IEM in Bangladesh. In addition, this study is the first report for the underlying genetic defects of the G6PD gene that causes G6PD deficiency in the Bangladeshi population and the establishment of a rapid and reliable HRM-based method for identification of G6PD variants among heterozygous females in Bangladesh.
Metabolomics Profiling for Amino Acidopathies of Infants in Hospital Based Settings in Bangladesh and Development of Easy-to-Use Kit for Disease Diagnosis and Monitoring
(©University of Dhaka, 2025-02-09) Hossain, Md. Shawkat
This study aimed to address the challenges associated with identifying inborn errors of metabolism (IEM) in resource-constrained environments, focusing on amino acid and galactose metabolism disorders in the Bangladeshi infant population. Liquid Chromatography Mass Spectrometry (LC-MS) is a robust diagnostic tool but faces limitations due to cost, complexity, and accessibility. Therefore alternative approaches, including enzyme assays, genetic testing, and metabolite profiling, were explored for effective IEM detection. In pursuit of an affordable diagnostic solution, a low-cost kit was developed for early detection and intervention of amino acid and galactose-related disorders. A validated HPLC-based method was established for accurate quantification of amino acids. The method exhibited strong linearity (R2 = 0.9999) and high accuracy (99.10% average recovery), establishing its suitability for amino acid analysis. Precision was confirmed through repeatability and intermediate precision tests, with low RSD values (0.30% to 1.53% and 0.23% to 0.66%, respectively). Amino acid cutoff ranges were determined for the Bangladeshi population using the validated HPLC method. Noteworthy variations in amino acid counts were observed in positive samples, emphasizing the importance of accurate cutoff values. A lateral flow paper-based diagnostic kit was designed for phenylalanine detection, offering a cost-effective means for phenylketonuria (PKU) monitoring. The kit demonstrated a minimum detection limit of 2 mg/dL, catering to PKU therapy needs. The study also concentrated on Galactosemia as a model disorder, underlining the necessity of early intervention. Molecular analysis of the GALT gene unveiled specific mutations in the Bangladeshi population, contributing to Galactosemia's molecular diversity. Evaluation of silent mutations' impact on splicing shed light on potential gene expression and protein function alterations, influencing Galactosemia outcomes. An easy-to-use lateral flow diagnostic kit for Galactosemia was developed, showing specificity for galactose and delivering rapid results within 5 minutes. Stability assessment revealed promising results at 2-8°C storage over a year, with signal degradation at higher temperatures, emphasizing proper storage conditions. The study highlights the potential of the HPLC method for amino acid detection, paving the way for accessible and cost-effective amino acidopathy detection facilities. The Galactosemia kit, with further refinement, holds promise for broader use. Insights into alternate GALT gene splicing patterns provide avenues for therapeutic exploration. Future research should fine-tune kits and establish precise phenylalanine and galactose cutoff ranges for the Bangladeshi population, contributing to improved IEM management.
Expression and regulation of efflux transporters in chemoresistant breast, colon and ovarian cancer cells in Bangladeshi patients
(© University of Dhaka, 2025-04-20) Deb Nath, Sudipta
Chemotherapy is a cornerstone in cancer treatment, but multidrug resistance (MDR) poses a substantial challenge. Hyperactivity of efflux pumps is one of the vital reasons to confer MDR. P-GP, MRP1 and ABCG2- these key ABC efflux pumps collectively transport a spectrum of chemotherapeutic drugs out of cells. HIF1A, a master regulator, is also associated with cancer biology, influencing cellular invasion, angiogenesis and metabolism. HuR regulates the expression of various carcinogenic molecules by mRNA trafficking, stabilization and enhancement of protein translation as well. Thereby this study addresses a crucial research gap in Bangladeshi population by inspecting the expression patterns of P-GP, MRP1 and ABCG2 in chemoresistant breast, colorectal and ovarian tissues. This study also aimed to delve into the mechanisms of drug resistance, focusing on HIF1A and HuR driven overexpression of ABC transporters, thus leading to chemoresistance. In this prospective study, 57 (chemoresistant) and 57 (paired control) tissue samples from Bangladeshi cancer patients were processed for immunofluorescence staining. The data showed the consistent co-expression of P-GP, MRP1 and ABCG2 across all cases, suggesting a potential synergistic function of transporter proteins in chemoresistance. The observations from this study also revealed that HIF1A and HuR overexpress in chemoresistant tissues, suggesting their pivotal role in chemoresistance mechanisms across malignancies and potential as therapeutic targets in Bangladeshi population. This study also suggests a direct regulation of P-GP, MRP1 and ABCG2 transcription by HIF1A that binds to the 'CGTG' sequence in the promoter regions. Besides, HuR might facilitate posttranscriptional mRNA stabilization by interacting with AU-rich elements (AREs) in the 3'UTR regions of the ABC transporters. Inhibition of these interactions of HIF1A and HuR appears as a promising approach to reverse chemoresistance. Thus, this study offers constructive insights for designing more efficient treatment strategies.
Impacts of pathogenic mutations and XmnI polymorphism on the severity of haemoglobin E β-thalassaemia patients in Bangladesh
(© University of Dhaka, 2025-07-07) Aziz, Md. Abdul
Background: Haemoglobin E β-thalassaemia is a common blood disorder in South Asia, especially Bangladesh. The disorder can range from mild and asymptomatic anemia to life-threatening anemia. Patients with asymptomatic anemia do not require blood transfusions in their lifetime. Mild Hb E beta thalassaemia patients require occasional blood transfusions. However, patients with life-threatening anemia need frequent blood transfusions. The presence of XmnI polymorphism with specific mutation led to a delay of blood transfusions, higher haemoglobin levels, better response to hydroxyurea treatment, and milder phenotypic presentation among Hb E β thalassaemia patients. Aims: The aim of this study is to examine how the HBB gene mutations and the XmnI polymorphism affect the severity and transfusion needs of haemoglobin E β thalassaemia patients. Methodology: In this study, a total of 360 Hb E β-thalassaemia patients were included. The seven common HBB mutations in Bangladesh were detected by amplification refractory mutation system (ARMS) method, and rare/novel mutations were detected by Sanger sequencing. Moreover, XmnI (PdmI) restriction site was detected by XmnI (PdmI) restriction enzyme digestion. Results: Among the subjects in this research, 62.5% (n=225) were regular transfusion dependent, 22.8% (n=82) were occasionally transfusion-dependent, 8.1% (n=29) were one-time transfusion patients, and 6.7% (n=24) were transfusion-independent patients. This study identified a total of thirteen pathogenic mutations among the 360 Hb E β thalassaemia patients. The most common mutation was IVS 1-5 (G>C), accounting for 57.8% (n=208) of cases, followed by Fr 41/42(-CTTT) 12.2% (n=44), codon 30 (G>C) 8.3% (n=30), Fr 8/9 (+G) 7.5% (n=27), codon 16 (-C) 3.9% (n=14), and codon 15 (G>A) 2.8% (n=10). The study also identified six rare mutations, including -90(C>T) 3.1% (n=11), IVS-I-130(G>A) 3.1% (n=11), IVS II-654 (C>T) 0.6% (n=2), and -30 (T>C) 0.3% (n=1), codon 14/15 (+G) 0.3% (n=1) and -29 (A>G) 0.3% (n=1). Of these, -29 (A>G) had not been reported previously in Bangladesh. Page | v This study found that those with codon 16 (-C) mutation required regular transfusions, while those with codon 30 (G>C) mutations had the least transfusion dependency (6.7%). Patients with codon 30 (G>C) mutations had the highest response to hydroxyurea therapy (76.7% (n=23)) (p=0.051), while patients with Codon 16 (-C) mutations had the lowest response (0% (n=14)) (p=0.057). Codon 16 (-C) mutation was associated with the least HbF (mean ± SD= 24.8±7.7%), while codon 30 (G>C) mutation was associated with the highest HbF (mean ± SD= 41.9±16.10%). Regarding XmnI polymorphism, Codon 16 (-C) mutation was associated with the highest wild type (-/-) (n=11) genotype (p=0.153), while codon 30 (G>C) mutation was associated with the highest homozygous mutated (+/+) (n=11) and the least wild type (-/-) (n=4) of XmnI polymorphism genotype (p=0.157). Conclusion: The outcomes of this study will be useful for developing treatment strategies involving HU therapy and for managing Hb E β-thalassemia patients in Bagladesh.
Molecular Basis of the Pelvi-Ureteric Junction Obstruction in Children with Hydronephrosis
(© University of Dhaka, 2025-11-05) SHADRUL ALAM, MOHAMMED
ABSTRACT: Background: Pelvi-Ureteric Junction Obstruction (PUJO) represents a significant cause of congenital hydronephrosis that can lead to progressive renal damage if untreated. While the condition is well-characterized clinically, its molecular pathogenesis remains poorly understood. This study aimed to identify key genetic alterations in PUJO through comprehensive transcriptomic analysis. Methods: We conducted a prospective study from January 2016 to December 2023 involving 9 pediatric patients (18 tissue samples) undergoing Anderson-Hynes pyeloplasty. Paired samples were collected from stenotic PUJ segments and adjacent normal ureteral tissue. RNA sequencing was performed followed by bioinformatic analysis to identify differentially expressed genes (DEGs) and pathway enrichment. Clinical parameters including demographics, presentation, and postoperative outcomes were systematically recorded. Results: The cohort showed male predominance (3.5:1 ratio) with mean age 40.22±32.61 months and left-sided predominance (77.8%). Key clinical presentations included abdominal distension (55.6%) and urinary tract infections (33.3%). Postoperative complications occurred in 22.2% cases. Transcriptomic analysis revealed 590 significant DEGs (292 upregulated, 298 downregulated), with notable enrichment in neuroactive ligand-receptor interactions (141 genes) and olfactory transduction pathways (50 genes). Chromosomal analysis identified 29 enriched regions, particularly on chromosomes 19, 17, and 9. Key dysregulated genes included KRT8 (upregulated) and SEMA3F (downregulated), implicating defects in smooth muscle differentiation and ureteral development. Conclusion: This first Bangladeshi transcriptomic study of PUJO identifies distinct molecular signatures involving neural signaling, ion transport, and developmental pathways. The findings provide novel insights into PUJO pathogenesis and potential biomarkers for early diagnosis. These results establish a foundation for developing targeted therapies to prevent renal damage in affected children.