Biology subjects

Adjaye, J.

Publications and source records attributed to Adjaye, J..

At least 19 recordsLinked to original sources

bioRxiv2026

Meta-analyses of the NEPTUNE dataset related to high and low risk FSGS and cellular APOL1 models identifies Calcium signalling, mTOR signaling and inflammation-associated pathways as operative in APOL1-mediated kidney disease.

BackgroundThe variants G1 and G2 within the APOL1 gene confer a higher risk of APOL1-mediated kidney disease (AMKD) whilst associated with an evolutionary advantage against trypanosome-mediated sleeping sickness. MethodsIn this study, we analysed transcriptome data of kidney biopsies from FSGS patients with the APOL1 high-risk (HR) and low-risk (LR) variants and compared it to cellular models based on patient-specific podocytes, HEK cells with engineered over-expressing HR variants and HR variant single-cell-RNA-seq data from kidney organoids. ResultsWe identified a signature of up- and down-regulated genes between biopsies from FSGS patients with APOL1 HR and LR variants. The up-regulated genes are functionally annotated to be associated with Calcium and mTOR signaling, whilst the down-regulated genes with inflammatory and immune response pathways. These pathways were confirmed by comparing with cellular models. Analysis of small molecules reverting the IFN-{gamma} stimulated gene expression to the non-stimulated gene expression in genome-edited APOL-G1 kidney organoids revealed several putative candidates such as the mTOR inhibitor AZD-2014. ConclusionWe have unveiled a signature of up- and down-regulated genes between APOL1 HR and LR kidney biopsies which could be assigned as associated with Calcium and mTOR signaling and down-regulated immune response.

molecular biology↗

bioRxiv2026

SAHA increases chaperone expression and reduces Z-alpha-1-antitrypsin polymers in a patient specific iPSC-based liver model for alpha-1-antitrypsin deficiency

The most severe phenotype of alpha-1-antitrypsin deficiency (AATD) is caused by the Z-mutation within the SERPINA1 gene. The Glu342Lys substitution causes misfolding and polymerisation of the alpha-1-antitrypsin (AAT) protein, its accumulation in the ER and increases the susceptibility of hepatocytes towards ER-stress. Here, we present an induced pluripotent stem cell (iPSC)-based hepatic model to study AATD. We demonstrate that iPSCs from AATD patients differentiate equally well to hepatocyte-like cells (HLCs) as control iPSCs. We detected ZAAT polymers in patient-derived HLCs which could be reduced by SAHA or CBZ treatment. Transcriptome analyses revealed major differences in metabolism and signalling between control and AATD HLCs and indicated increased stress levels affecting intracellular organelles. Importantly, the transcriptomes of control and patient-derived cells separated into distinct clusters with respect to the expression of Heat-shock protein (HSP) encoding genes. SAHA treatment increased expression of various HSPs which might contribute towards reduced ZAAT polymers.

cell biology↗

bioRxiv2025

Human CSB-deficient iPSCs exhibit impaired DNA damage repair and stress responses following BPDE exposure in an early developmental model.

Maintenance of genome integrity is essential for normal human development, particularly during pre-gastrulation stages when rapid proliferation and intense transcriptional activity increase susceptibility to DNA damage. Environmental genotoxins such as benzo[a]pyrene (BaP), a widespread polycyclic aromatic hydrocarbon, and its reactive metabolite benzo[a]pyrene diol epoxide (BPDE) form bulky DNA adducts that interfere with replication and transcription, thereby posing significant risks to embryonic genome stability. To examine how genetic defects in DNA repair influence these effects, we assessed human induced pluripotent stem cells (iPSCs) carrying pathogenic mutations in ERCC6 (encoding the Cockayne syndrome B, CSB, protein), a key component of transcription-coupled nucleotide excision repair. Pathogenic ERCC6 mutations result in Cockayne syndrome- a severe neurodevelopmental disorder characterized by growth failure, premature aging, and multisystemic degeneration, thus underscoring the essential developmental functions of CSB. Exposure of healthy and CSB-deficient patient derived iPSCs to BPDE revealed impaired proliferation, persistent accumulation of DNA damage and defective checkpoint activation in CSB-deficient lines. Although the levels of key pluripotency-regulating proteins such as OCT4 and NANOG remained unaltered, we observed altered levels of SOX2 and p-SMAD1/5 signaling thus implying that unrepaired DNA damage can perturb developmental-associated signaling pathways and biological processes. Transcriptomic profiling revealed broad suppression of DNA repair and cell-cycle pathways together with activation of p53-, TNF-, and MAPK/JNK-mediated stress responses in CSB-deficient lines. Failure to induce anti-oxidant defenses, including SOD2 and IDO, further contributed to oxidative imbalance and incomplete apoptotic clearance. These findings demonstrate that CSB function is essential for coupling DNA repair with transcriptional recovery and redox homeostasis in pluripotent cells. Loss of CSB destabilizes the genome stability under genotoxic stress, providing a mechanistic basis for developmental toxicity of environmental polycyclic aromatic hydrocarbons and underscoring the importance of considering genetic susceptibility in developmental toxicology risk assessment.

developmental biology↗

bioRxiv2025

Urinary proteins from Sickle Cell patients induce inflammation and kidney injury via the TGFβ-p53 axis in a podocyte cell culture model.

BackgroundSickle cell disease (SCD) is an inherited blood disorder affecting the oxygen-carrying hemoglobin in red blood cells making them deform into a sickle shape. Hemolysis and vaso-occlusion associated with this process can lead to complications in many organs and frequently to renal complications. Numerous factors are considered to contribute towards the development of proteinuria (PU) in SCD including hyperfiltration, ischemia, oxidative stress and decreased nitric oxide (NO) bioavailability but the detailed pathophysiology still needs further elucidation. MethodsEmploying arrays, we investigated cytokines and kidney injury-associated markers in the urine of a cohort of SCD patients from Ghana carrying the SS and SC genotypes which were further sub-divided into groups with proteinuria (SCD_PU) and without proteinuria (SCD). ResultsWe identified up-and down-regulated proteins when comparing SCD with and without proteinuria. Amongst these is the well-established kidney injury marker-Clusterin which was up-regulated and could be validated in an ELISA-based assay. Refining the study to the SS and SC genotypes, we identified (and confirmed by ELISA) another established kidney injury marker-NGAL, as up-regulated in both genotypes and SCD with and without proteinuria. Metascape-based analysis of biological processes revealed "Cellular component disassembly" associated with proteins expressed in SCD but not regulated between PU and no PU and "leukocyte chemotaxis" down-regulated in SCD_PU vs. SCD. Interestingly, "Integrin-cell-surface interactions" was associated with proteins up-regulated between SCD_PU vs. SCD which is consistent with endothelial hyperplasia in the setting of glomerular hyperfiltration. To investigate the effect secreted urine proteins have on human podocytes in vitro, immortalized podocytes supplemented with SCD_PU urine showed elevated p53 levels in both immunofluorescence staining and RT-PCR compared to SCD. Additionally, RT-PCR revealed elevated levels of VEGF, NGAL and the pro-inflammatory proteins-TGF{beta}, IL6, IL8 and TNF. ConclusionWe hypothesize that the increased number of endothelial cells in hyperplasia and hyperfiltration leads to more Integrin-mediated links to podocyte foot processes at the glomerular basement membrane and to glomerular fibrosis. Severe inflammation and kidney injury in SCD_PU patients is induced by the TGF{beta}-p53 axis.

molecular biology↗

bioRxiv2025

Modelling APOL1-mediated kidney inflammation and fibrosis using a partially reprogrammed urine derived SIX2-positive renal progenitor cell line.

BackgroundCKD affects approximately 850 million people worldwide and is a leading cause of mortality. Podocytes, cells in the kidney are terminally differentiated and incapable of division in vivo making the establishment of primary cultures particularly challenging. The ability of cells to proliferate and avoid senescence is closely linked to telomere length. When telomere length becomes critically reduced, it results in cellular senescence. MethodsWe present the successful rejuvenation of a human SIX2-positive renal progenitor cell line derived from the urine of a 30-year-old West African male (UM30-OSN). To achieve partial reprogramming, plasmids expressing the Yamanaka factors OCT4, SOX2, NANOG, c-Myc, and KLF4 were employed. ResultsUM30-OSN expresses the pluripotency-associated marker SSEA4, renal stem cell markers such as SIX2, CD133 and CD24, determined by immunofluorescence, FACS and qPCR. Expression analysis revealed downregulation of senescence markers p21and p53 and upregulation of proliferation-associated genes PCNA, KI67 and TERT, confirming rejuvenation. Upon podocyte differentiation, UM30-OSN cells expressed podocyte-specific markers NPHS1, NPHS2, SYNPO and CD2AP. Comparative transcriptome analyses revealed a correlation co-efficiency (R2 = 0.88) with the immortal podocyte line AB 8/13. To demonstrate the usefulness of UM30-OSN to model APOL1-mediated kidney disease, we investigated the effects of Interferon-{gamma} (IFN-{gamma}) on UM30-OSN derived podocytes and evaluated the potential of the JAK1/JAK2 inhibitor Baricitinib to mitigate IFN-{gamma}-induced cellular responses. IFN-{gamma} stimulation resulting in increased phosphorylation of STAT1, activation of APOL1, upregulation of pro-inflammatory and fibrotic markers such as, IL-6, TGF-{beta}, Vimentin, Fibronectin, and morphological changes indicative of cell stress. Pre-treatment with Baricitinib effectively inhibited STAT1 phosphorylation, reduced expression of pro-inflammatory and fibrosis-associated genes, and preserved podocyte morphology. ConclusionGiven their robust proliferation capacity, UM30-OSN cells represent a valuable additional model for investigating kidney-associated diseases such the contribution of APOL1 high-risk variants to kidney injury and fibrosis.

molecular biology↗

bioRxiv2025

Human Urine-derived SIX2-positive renal progenitor cells improve kidney injury in an IRI mouse model

Cells with the characteristics of renal progenitor cells are shed in urine. The aim of this study was to investigate whether these SIX2-positive urine-derived renal progenitor cells (UdRPC) have therapeutic potential in treating or managing acute to chronic kidney injuries. Human UdRPC were obtained from a woman aged 35 years, expanded, and characterized in vitro and finally transplanted unilaterally under the renal capsule of mouse kidneys that had undergone ischemia reperfusion injury (IRI). Analyses of blood serum from the mice revealed that the transplanted human UdRPC transiently influenced the secretome of the animals by triggering the release of inflammatory and angiogenic factors. Of obvious therapeutic relevance, however, was the finding that the transplanted cells suppressed renal fibrosis resulting from IRI. The expression of genes linked to inflammation and chronic kidney disease was also reduced in mouse kidneys in the presence of UdRPC. Ultimately, the human UdRPC were able to alleviate the severity of the injury through the amelioration of fibrosis but were unable to restore complete kidney function within 21 days. Nevertheless, the findings of this study demonstrate a significant influence of transplanted UdRPC on kidney fibrosis after acute to chronic injury presumably via an altered secretome.

cell biology↗

bioRxiv2025

Human blood vessel organoids recapitulate key mechanisms of transition from vasculopathy to fibrosis in systemic sclerosis

Systemic sclerosis (SSc) is an autoimmune disease that transitions from vasculopathy as an initiating pathogenic event to tissue fibrosis. The mechanisms of these transitions remain, however, poorly understood, mainly because complex multicellular human models of SSc vasculopathy are lacking. Here we characterized blood vessel organoids (BVOs) as a novel model system of vasculopathy in SSc. We demonstrate that exposure of SSc-BVOs to SSc serum triggers changes on epigenetic, mRNA and protein levels and recapitulates key pathogenic features of SSc vasculopathy, with shifts from angiogenic endothelial cell subsets to those undergoing endothelial-to-mesenchymal transition, loss of endothelial cells-pericytes interactions and profound angiogenic defects. The genetic predisposition of SSc donors and serum IgGs are required for the deleterious effects of SSc serum. We further validate SSc-BVOs as a human model system to evaluate candidate therapies targeting SSc microvasculopathy and use this system to provide evidence that {gamma}-secretase inhibition is a potential therapeutic approach.

biophysics↗

bioRxiv2025

Evidence for conserved gene expression and biological processes operative in human podocytes and adult brain

BackgroundPodocytes are essential for the proper functioning of the glomerular filtration barrier and are characterized by their intricate structure which includes well-organized primary and secondary foot processes. The brain expresses podocyte-associated proteins such as Nephrin, and Synaptopodin which are well documented. Processing of information and inter-cellular communication between podocytes and neurons employ similar molecular mechanisms which include actin-based projections, adhesion molecules, and signaling pathways. MethodsIn this study, we analysed brain-associated biological processes, transcription factors and pathways significantly regulated in UdPodocytes, i.e. podocytes differentiated from SIX2-positive UdRPCs (urine-derived renal progenitor cells). We compared gene expression in iPSC-derived brain and kidney organoids with UdPodocytes and mapped the overlapping 344 genes to brain regions via the GTEX (Genotype-Tissue Expression) database and investigated their regulation induced by the mediator of the renin-angiotensin system-angiotensin II (ANG II) ResultsThe protein interaction network of UdPodocytes genes associated with brain in GTEX contains modules for pre-synapse, post-synapse, endocrine processes and neural crest differentiation. We also found that the genes overlapping between brain and podocytes are also expressed in iPSC-derived kidney and brain organoids and could map the involved genes to all regions of the brain with the frontal cortex as the most enriched. The genes SUZ12, NFKB1 and PAX2 were the most significantly over-represented transcription factors. We independently confirmed the conserved expression of PAX6, KCNQ3, TUJ1,MAP2, TAU and ACTN4 in Udpodocytes as shown by Western blotting, Immunofluorescence and RT-PCR. We further investigated if the conserved genes are also regulated by ANGII. This unveiled genes down-regulated upon ANGII-stimulation of Udpodocytes to be associated with axon guidance, Calcium, Hippo and cGMP-PKG signaling as over-represented pathways. ConclusionIn conclusion, we have identified 344 genes with overlapping expression in brain and podocytes. These are mainly associated with synaptic signaling and cell projections. This implies that human urine-derived SIX2-renal progenitor cells differentiated into podocytes can serve as a platform for dissecting and understanding the relevance of conserved biological processes in podocytes which are currently annotated as neuron projection, axons, neurogenesis and synaptic signaling.

molecular biology↗

bioRxiv2025

Impact of nephrotoxins and oxidants on survival and transport function of hiPSC-derived renal proximal tubular cells

Due to their role in excretion, renal proximal tubule cells are susceptible to damage by toxic metabolites and xenobiotics. The regenerative capacity of the kidney allows for the replacement of damaged cells, a process involving differentiation programs. However, kidney function tends to decline, suggesting that the replacement cells may not achieve full functionality. To understand possible causes of this decline, we investigated effects of nephrotoxins and oxidants on the differentiation of induced pluripotent stem cells (iPSC) into proximal tubular epithelial-like cells (PTELC). Proliferation, apoptosis, senescence and expression of oxidative defense genes were analyzed in iPSCs, differentiating and differentiated cells treated with cisplatin (CisPt, up to 45 {micro}M), cyclosporin A (CycA, up to 12 {micro}M) and the oxidants menadione (Mena, up to 50 {micro}M) and tert-butylhydroquinone (tBHQ, up to 50 {micro}M). We found that differentiating cells were most sensitive to oxidants and showed increased sensitivity to CisPt, whereas all differentiation stages showed similar sensitivity to CycA. Both oxidative stress and CisPt triggered apoptosis in all differentiation stages, whereas CycA mainly induced senescence. Treatment during differentiation resulted in long-term effects on gene expression in differentiated cells. While oxidants had no effect on transport function of differentiated cells, CisPt and CycA impaired albumin uptake. Our data suggest a substantial sensitivity of differentiating cells to nephrotoxins and oxidants, an aspect that could potentially interfere with regenerative processes.

pharmacology and toxicology↗

bioRxiv2024

Investigating BPDE-induced embryonic toxicity employing hiPSC-based models

Benzo[a]pyrene diol epoxide (BPDE) is a metabolite of the environmental contaminant Benzo[a]pyrene- a byproduct of incomplete combustion of organic matter. BPDE reacts with DNA to form BPDE-DNA bulky adducts which if not removed can lead to mutations due to DNA base-pair substitutions. While the effects of BPDE on somatic cells are fairly well described, its effects on early human development are currently unknown. In this study, we investigated for the first time the effect of BPDE on human induced pluripotent stem cells (hiPSCs) and their differentiated neuroprogenitor cells (NPCs) as a model for early embryonic development. Furthermore, we compared hiPSCs and NPCs derived from cells of patients suffering from Nijmegen Breakage Syndrome (NBS), which is a chromosomal instability disorder characterized by defective DNA repair and increased risk of malignancies. Transcriptome analysis, coupled with protein content analysis employing immunostaining and Western blots, revealed that hiPSCs are more sensitive to BPDE exposure when compared to NPCs with an enhanced expression of several genes associated with p53-mediated DNA damage response, including DNA repair by lesion bypass, cell cycle checkpoints and extrinsic apoptosis. We also identified that cells from NBS patients showed less apoptotic response and a distinct p53 response than their healthy counterparts. This iPSC-based study enhances our meagre knowledge of the effects of BPDE on early human development in both healthy individuals and NBS patients. Furthermore, our model conforms with the 3Rs principle.

pharmacology and toxicology↗

bioRxiv2024

Inhibiting DPP4 activity protects hiPSC-derived steatotic HLCs by supporting fatty acid and purine metabolism and dampening inflammation

BackgroundMetabolic dysfunction-associated steatotic liver disease (MAFLD) has a high prevalence and high co-morbidity for other diseases. Due to the complexity of this multifactorial disease, therapy options are still rather limited. We employed an in vitro pluripotent stem cell-based model to decipher potential disease-associated molecular pathways and to study the mode of action of prospective drugs. Dipeptidyl peptidase 4 (DPP4) or Cluster of differentiation 26 (CD26) is involved in inflammation, infections, immune disorders, type 2 diabetes, kidney disease and cancer. MethodsWe induced the steatosis phenotype in human induced pluripotent stem cell (iPSC) derived hepatocyte-like cells (HLCs) by oleic acid (OA)-feeding and confirmed regulation of clinically relevant pathways by NGS-based global transcriptomic analyses. Analysis of the secretome of steatotic HLCs revealed DPP4 as a potential key mediator of the disease. To further elucidate its role in the development of MAFLD, we inhibited DPP4 activity with Vildagliptin (VILDA) and analyzed the global transcriptome changes as well as specific gene and protein expression of steatosis-associated genes with and without DPP4 inhibition. ResultsMAFLD-associated pathways such as PPAR- and TNF signaling were differentially regulated in hiPSC-derived steatotic HLCs. We found increased hepatic DPP4 activity and secretion upon OA. Gene expression of fatty acid and purine metabolism and inflammation-associated pathways were regulated upon DPP4 inhibition. ConclusionsOur HLC-model confirmed association of DPP4 with metabolism and inflammation which foster the development of MAFLD. Inhibiting DPP4 with VILDA partially relieved the steatotic phenotype on a global transcriptomic level. Impact and implicationsGiven the difficulties of identifying suitable anti-MAFLD drugs, novel model systems are urgently needed. Our in vitro HLC-model reproduced DPP4-dependent aspects of the disease and responded positively to Vildagliptin treatment. Further elucidation of the role of DPP4 in the etiology of MAFLD and other diseases is warranted.

cell biology↗

bioRxiv2024

Single cell data enables dissecting cell types present in bulk transcriptome data

The quality of organoid models can be assessed by single-cell-RNA-sequencing (scRNA-seq) but often only bulk transcriptome data is available. Here we present a pipeline for the analysis of scRNA-seq data and subsequent "deconvolution", which is a method for estimating cell type fractions in bulk transcriptome data based on expression profiles and cell types found in scRNA-seq data derived from biopsies. We applied this pipeline on bulk iPSC-derived kidney and brain organoid transcriptome data to identify cell types employing two scRNA-seq kidney datasets and one brain dataset. Relevant cells present in kidney (eg. proximal tubules, distal convoluted tubules and podocytes) and brain (eg. neurons, astrocytes, oligodendrocytes and microglia) with obligatory endothelial and immune-related cells were identified. We anticipate that this pipeline will also enable estimation of cell type fractions in organoids of other tissues.

bioinformatics↗

bioRxiv2024

Identification of novel evolutionarily conserved genes and pathways in human and mouse musculoskeletal progenitors

The axial skeletal system and skeletal muscles of the vertebrates arise from somites, the blocks of tissues flanking both sides of the neural tube. The progenitors of Somites, called the Presomitic Mesoderm (PSM) reside at the posterior end of a developing embryo. Most of our understanding about these two early developmental stages comes from the studies on chick and mouse, and in the recent past, there have been a few studies on human. Here, we have analysed and compared the RNA-sequencing data of PSM and somite tissues from Mouse and Human. The functional and pathway enrichment analysis identified the key Hub-genes that are evolutionarily conserved in the PSM and the somites of both the organisms that include 23 multifunctional genes likely to be associated with different developmental disorders in humans. Our analysis revealed that NOTCH, WNT, MAPK, BMP, Calcium, ErbB, cGMP-PKG, RAS and RAP1 signaling pathways are conserved in both human and mouse during the development of PSM and Somites. Furthermore, we validated the expression of representative conserved candidates in the hESCs-derived PSM and somite cells (NOG, BMP2, BMP7, BMP5, HES5 and MEF2C). Taken together, our study identifies putative gene interactions and pathways that are conserved across the mouse and human genomes, which may potentially have crucial roles in human PSM and somite development.

bioinformatics↗

bioRxiv2024

Hemozoin induces Malaria via activation of DNA damage, p38 MAPK and Neurodegenerative Pathways in Human iPSC-derived Neuronal Model of Cerebral Malaria

Malaria caused by Plasmodium falciparum infection results in severe complications including cerebral malaria (CM), in which approximately 30% of patients end up with neurological sequelae. Sparse in vitro cell culture-based experimental models which recapitulate the molecular basis of CM in humans has impeded progress in our understanding of its etiology. This study employed healthy human induced pluripotent stem cells (iPSCs) derived neuronal cultures stimulated with hemozoin (HMZ)-the malarial toxin as a model for CM. Secretome, qRT-PCR, Metascape, and KEGG pathway analyses were conducted to assess elevated proteins, genes, and pathways. Neuronal cultures treated with HMZ showed enhanced secretion of interferon-gamma (IFN-{gamma}), interleukin (IL)1-beta (IL-1{beta}), IL-8 and IL-16. Enrichment analysis revealed malaria, positive regulation of cytokine production and positive regulation of mitogen-activated protein kinase (MAPK) cascade which confirm inflammatory response to HMZ exposure. KEGG assessment revealed up-regulation of malaria, MAPK and neurodegenerative diseases-associated pathways which corroborates findings from previous studies. Additionally, HMZ induced DNA damage in neurons. This study has unveiled that exposure of neuronal cultures to HMZ, activates molecules and pathways similar to that observed in CM and neurodegenerative diseases. Furthermore, our model is an alternative to rodent experimental models of CM.

neuroscience↗

bioRxiv2023

Cockayne syndrome patient iPSC-derived brain organoids and neurospheres show early transcriptional dysregulation of biological processes associated with brain development and metabolism.

Cockayne syndrome is a rare hereditary autosomal recessive disorder characterized by diverse neurological afflictions. However, little is known about the cerebral development in CS patients. We generated neurospheres and cerebral organoids utilizing Cockayne Syndrome B Protein (CSB) deficient induced pluripotent stem cells derived from two patients with distinct severity levels of CS and healthy controls. The transcriptome of both developmental timepoints was explored using RNA-Seq and bioinformatic analysis to identify dysregulated biological processes common to both CS patients in comparison to control. CSB-deficient neurospheres displayed upregulation of VEGFA-VEGFR2 signaling pathway, Vesicle-Mediated transport and head development. CSB-deficient cerebral organoids exhibited downregulation of brain development, neuron projection development and synaptic signalling. We further identified upregulation of Steroid Biosynthesis as common to both timepoints, in particular upregulation of the Cholesterol Biosynthesis branch. Our results provide insights into the neurodevelopmental dysregulation in CS patients and strengthen the theory, that CS is not only a neurodegenerative, but also a neurodevelopmental disorder.

neuroscience↗

bioRxiv2023

Unconjugated bilirubin induces neuro-inflammation in an induced pluripotent stem cell-derived cortical organoid model of Crigler Najjar Syndrome

Bilirubin-induced neurological damage (BIND), which is also known as Kernicterus, occurs as a consequence of defects in the bilirubin conjugation machinery, thus resulting in unconjugated bilirubin (UCB) to cross the blood-brain barrier (BBB) and accumulation. Severe hyperbilirubinemia can be caused by a mutation within the UGT1A1 encoding gene. This mutation has a direct contribution towards bilirubin conjugation leading to Kernicterus as a symptom of Crigler Najjar Syndromes (CNS1, CNS2) and Gilbert syndrome, which results in permanent neurological sequelae. In this comparative study, we used human induced pluripotent stem cells (hiPSCs)-derived 3D-brain organoids to model BIND in vitro and unveil the molecular basis of the detrimental effects of UCB in the developing human brain. hiPSC-derived from healthy and CNS patients were differentiated into day-20 brain organoids, these were then stimulated with 200nM UCB. Analyses at 24- and 72-hrs post-treatment point at UCB-induced neuro-inflammation in both cell lines. Transcriptome and associated KEGG and Gene Ontology analyses unveiled activation of distinct inflammatory pathways such as cytokine-cytokine receptor interaction, MAPK signaling, calcium signaling, NF{kappa}B activation. Furthermore, both mRNA expression and secretome analysis confirmed an upregulation of pro-inflammatory cytokines such as IL6 and IL8 upon UCB stimulation. In summary, this novel study has provided insights into how a human iPSC-derived 3D-brain organoid model can serve as a prospective platform for studying the etiology of BIND-Kernicterus.

neuroscience↗

bioRxiv2023

Cytokines in the Urine of AKI patients regulate TP53 and SIRT1 and can be used as biomarkers for the early detection of AKI

Acute kidney injury (AKI) is a major kidney disease with a poor clinical outcome. It is a common complication with an incidence of 10-15% of patients admitted to hospital. This rate even increases for patients who are admitted to the intensive care unit with an incidence of >50%. AKI is characterized by a rapid increase in serum creatinine, decrease in urine output, or both. Associated Symptoms include feeling sick or being sick, diarrhoea, dehydration, decreased urine output-although occasionally urine output remains normal, fluid retention-causing swelling in the legs or ankles, shortness of breath, fatigue and nausea. However, sometimes acute kidney injury causes no signs or symptoms and is detected by lab tests. Therefore, an urgent demand for non-invasive biomarkers for early detection of AKI are highly desirable. This might enable the prevention of the progression from AKI to CKD. In this study, we analysed the secretome of urine of an AKI patient cohort employing a kidney-biomarker cytokine assay. Based on these results we suggest, ADIPOQ, EGF and SERPIN3A as potential biomarkers, which might be able to detect AKI as soon as 24 h post-surgery. For the later stages, common biomarkers for the detection of AKI in both male and female patients we suggest, VEGF, SERPIN3A, TNFSF12, ANPEP, CXCL1, REN, CLU and PLAU. These markers in combination might present a robust strategy to identify the development of AKI as early as 24h or 72h post-surgery. Furthermore, we evaluated the effect of patient and healthy urine on human podocyte cells. We conclude that cytokines in the urine of AKI patients trigger processes which are needed to repair the damaged nephron and activate TP53 and SIRT1 to maintain the balance between proliferation, angiogenesis, and cell cycle arrest. In conclusion, the Renin-Angiotensin pathway seems to have major implications.

molecular biology↗

bioRxiv2022

Urine-based detection of biomarkers indicative of chronic kidney disease in a patient cohort from Ghana

Chronic kidney disease (CKD) is a global health burden with a continuously increasing prevalence associated with an increasing incidence of diabetes and hypertension in aging populations. The CKD definition of a more than three months lasting low glomerular filtration rate (GFR) or other renal impairments including proteinuria implies that multiple factors may contribute to the disease. While there are indications of ethnic differences it is hard to disentangle these from confounding social factors. Usually, CKD is detected in later stages of the disease when irreversible renal damage has already occurred, thus suggesting a need for early non-invasive diagnostic markers. In this study, we explored the urine secretome of a CKD patient cohort from Ghana employing a kidney-injury and a more general cytokine assay. We identified panels of kidney-specific cytokine markers which were also gender-specific and a panel of gender-independent cytokine markers. The gender-specific markers are IL10 and MME for male and CLU, RETN, AGER, EGFR and VEGFA for female. The gender-independent cytokine markers were APOA1, ANGPT2, C5, CFD, GH1, ICAM1, IGFBP2, IL8, KLK4, MMP9 and SPP1 (up-regulated) and FLT3LG, CSF1, PDGFA, RETN and VEGFA (down-regulated). APOA1 - the major component of HDL particles - was up-regulated in Ghanaian CKD patients and its co-occurrence with APOL1 in a subpopulation of HDL particles may point to specific CKD-predisposing APOL1 haplotypes in patients of African descent - this however needs further investigation. The identified panels may lay down the foundation for CKD-biomarker assays to be confirmed in further studies with a larger cohort of patients.

molecular biology↗