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Zhao, S.

Publications and source records attributed to Zhao, S..

At least 109 records · Page 6Linked to original sources
bioRxiv2025

Detection of Diffuse to Focal Myocardial Fibrosis by Cardiovascular Magnetic Resonance Against Histology in Mini-Swine

BackgroundMyocardial fibrosis predicts adverse outcomes in myocardial infarction (MI) and other cardiovascular conditions. Cardiovascular magnetic resonance (CMR) can non-invasively detect focal and diffuse myocardial fibrosis, but comprehensive validation against histology remains scarce. AimsTo assess the comparative diagnostic performance of CMR methods in detecting myocardial fibrosis, using a mini-swine model and histology as gold standard. MethodsEighteen mini-swine (16 MI; 2 healthy) underwent CMR cine, LGE, T1- and ECV-mapping. Two commonly-used T1-mapping methods - MOLLI 5(3)3 and ShMOLLI 5(1)1(1)1 - were included. Pathological sections were categorized as infarcted, peri-infarct, remote and healthy myocardium based on triphenyl tetrazolium chloride staining. Fibrotic burden was quantified by collagen volume fraction (CVF) into severe (CVF[&ge;]30%), moderate (CVF=10-25%), and mild (CVF=3-14%). The relationships between LGE, T1, ECV and CVF, and diagnostic performance using area-under-the-curve (AUC), were analyzed. ResultsFor detecting severe fibrosis, LGE, T1 and ECV all had excellent diagnostic performance (AUC: LGE=0.93, ECVShMOLLI=0.96, T1ShMOLLI=0.91, T1MOLLI=0.93, ECVMOLLI=0.88). ECVShMOLLI showed significantly better discriminatory accuracy than ECVMOLLI in detecting severe fibrosis and MI (both p<0.05), with the highest correlation to CVF (ECVShMOLLI r=0.86, ECVMOLLI r=0.82, T1ShMOLLI r=0.77, T1MOLLI r=0.77, semi-quantitative LGE r=0.75). Only T1ShMOLLI and ECVShMOLLI, but not LGE or T1MOLLI/ECVMOLLI, differentiated remote myocardium with mild fibrosis (CVF=8.23%) from healthy myocardium (CVF=2.01%). ConclusionsCMR can detect severe to mild myocardial fibrosis as validated against histology. For low-grade fibrosis, T1-mapping significantly outperformed LGE. Choice of CMR methodologies matters for myocardial fibrosis detection, which has importance for clinical trial design.

pathology↗
bioRxiv2025

Development of FAP-targeted theranostics discovered by next-generation sequencing-augmented mining of a novel immunized VNAR library

Cancer-associated fibroblasts (CAFs) in the stroma of solid tumors promote an immunosuppressive tumor microenvironment (TME) that drives resistance to therapies. The expression of the protease fibroblast activation protein (FAP) on the surface of CAFs has made FAP a target for development of therapies to dampen immunosuppression. Relatively few biologics have been developed for FAP and none have been developed that exploit the unique engagement properties of Variable New Antigen Receptors (VNARs) from shark antibodies. As the smallest binding domain in nature, VNARs cleverage unique geometries and recognize epitopes conventional antibodies cannot. By directly immunizing a nurse shark with FAP, we created a large anti-FAP VNAR phage display library. This library allowed us to identify a suite of anti-FAP VNARs through traditional biopanning and also by an in silico approach that did not require any prior affinity-based enrichment in vitro. We investigated four VNAR-Fc fusion proteins for theranostic properties and found that all four recognized FAP with high affinity and were rapidly internalized by FAP-positive cells. As a result, the VNAR-Fc constructs were effective antibody-drug conjugates in vitro and were able to localize to FAP-positive xenografts in vivo. Our findings establish VNAR-Fc constructs as a versatile platform for theranostic development that could yield innovative cancer therapies targeting the TME.

synthetic biology↗
bioRxiv2024

Integrated analysis of COVID-19 multi-omics data for eQTLs reveals genetic mechanisms underlying disease severity

The global pandemic caused by the SARS-CoV-2 virus provided an unprecedented opportunity to investigate genetic factors influencing the disease severity of the viral infection. Despite a plethora of recent research on both SARS-CoV-2 and COVID-19, few have taken a systems biology approach to address individual-level variation, especially based on non-European populations. Accordingly, we analyzed multi-omics data generated at three timepoints from 193 Korean COVID-19 patients with mild or severe symptoms, composed of whole genome sequencing, blood-based single-cell RNA-sequencing (2.15M cells), 195 cytokine profiles, and human leukocyte antigen (HLA) allele data. We identified expression quantitative trait loci (eQTLs), disease severity interacting eQTLs (n = 388), and disease progression interacting eQTLs (n = 945) for various cell types. We elucidated a complex regulatory mechanism involving HLA genes and their targets, and identified genetic determinants of cytokine levels. Finally, we show how regulation of ieQTLs is established by upstream transcription factors (TFs), illustrating complex regulation of the IGFBP7 ieQTL by a combined action of two TFs, which is potentially important in conferring differential severity. This study illuminates an efficient molecular interrogation framework that can be applied toward understanding infectious disease progression in individuals of different genotypes.

genetics↗
bioRxiv2024

Functional fingerprinting for the developing brain using deep metric learning: an ABCD study

Brain fingerprinting is a promising approach for characterizing the uniqueness of individual brain functioning using functional magnetic resonance imaging (fMRI) data. Here, we propose a novel deep learning framework, the metric-BoIT, for brain fingerprinting and demonstrate its effectiveness in capturing individual variability among early adolescents undergoing dramatic brain changes. Utilizing resting-state fMRI data from the Adolescent Brain Cognitive Development (ABCD) dataset, we identified brain functional fingerprints that achieved remarkable individual identification accuracy, reaching 97.6% within a single session and maintaining 86.6% accuracy over a four-year developmental period. Annotation analysis revealed that higher-order association regions, particularly those within the default-mode network, contributed most significantly to these distinctive brain fingerprints (t = 5.618, p < 0.001). Moreover, these brain fingerprints were relevant to cognitive functions, as evidenced by significant correlations with fluid intelligence (F = 1.282, p = 0.027) and crystallized intelligence (F = 1.405, p < 0.001). The extracted brain fingerprints were additionally associated with genetics, showing that individuals with strong genomic relationships exhibited more similar brain fingerprint patterns (t = -12.330, p < 0.001). Together, our study not only presents an innovative approach to brain functional fingerprinting but also provides valuable insights into the individual variability underlying adolescent neurodevelopment.

neuroscience↗
bioRxiv2024

RNA-programmable cell type monitoring and manipulation in the human cortex with CellREADR

Reliable and systematic access to diverse cell types is necessary for understanding the organization, function, and pathophysiology of human neural circuits. Methods for targeting human neural populations are scarce and currently center on identifying transcriptional enhancers and engineering viral capsids. Here we demonstrate the utility of CellREADR, a programmable RNA sensor-effector technology that couples cellular RNA sensing to effector protein translation, for accessing, monitoring, and manipulating specific neuron types in human cortex, ex vivo. We designed CellREADRs to target two subpopulations, CALB2 GABAergic interneurons and FOXP2 glutamatergic projection neurons, then validated targeting specificity using histological, electrophysiological, and transcriptomic methods. CellREADR expression of channelrhodopsin and GCamp enabled the manipulation and monitoring of these populations in live cortical microcircuits. By demonstrating specific, reliable, and programmable experimental access to human neuronal subpopulations, our results highlight CellREADRs potential for studying neural circuits and treating brain disorders.

neuroscience↗
bioRxiv2024

RNA programmable cell targeting and manipulation with CellREADR

Methods that provide specific, easy, and scalable experimental access to animal cell types and cell states will have broad applications in biology and medicine. CellREADR - Cell access through RNA sensing by Endogenous ADAR (adenosine deaminase acting on RNA), is a programmable RNA sensor-actuator technology that couples the detection of a cell-defining RNA to the translation of an effector protein to monitor and manipulate the cell. The CellREADR RNA device consists of a 5 sensor region complementary to a cellular RNA and a 3 payload coding region; payload translation is gated by the removal of a STOP codon in the sensor region upon base pairing with the cognate cellular RNA through an ADAR-mediated A- to-I editing mechanism ubiquitous to metazoan cells. CellREADR thus highlights the potential for RNA-based monitoring and manipulation of animal cells in ways that are simple, versatile, and generalizable across tissues and species. Here, we describe a detailed protocol for implementing CellREADR experiments in cell cultures and in animals. The procedure includes sensor and payload design, cloning, validation and characterization in mammalian cell cultures. The in vivo animal protocol focuses on AAV-based delivery of CellREADR using brain tissue as examples. We describe current best practices, various experimental controls and trouble-shooting steps. Beginning from sensor design, the validation of RNA sensor-actuators in cell cultures can be completed in 2-3 weeks. The construction of AAV vectors and their in vivo validation and characterization will take additional 6-8 weeks.

molecular biology↗
bioRxiv2024

Targeting SLC7A11-mediated cysteine metabolism for the treatment of trastuzumab resistant HER2 positive breast cancer

Trastuzumab resistance remains a challenge for HER2 positive breast cancer treatment. Targeting metabolic reprogramming would provide novel insights for therapeutic strategies. Here, we integrated metabolomics, transcriptomics and epigenomics data of trastuzumab sensitive and primary resistant HER2 positive breast cancer to identify metabolic alterations. Aberrant cysteine metabolism was discovered in trastuzumab primary resistant breast cancer at both circulating and intracellular levels. The inhibition of SLC7A11 and cysteine starvation could synergize with trastuzumab to induce ferroptosis. Mechanistically, increased H3K4me3 and decreased DNA methylation enhanced SLC7A11 transcription and cystine uptake in trastuzumab resistant breast cancer. The regulation of epigenetic modifications modulated cysteine metabolism and ferroptosis sensitivity. These results revealed an innovative approach for overcoming trastuzumab resistance by targeting specific amino acid metabolism.

cancer biology↗
bioRxiv2024

Glycophagy is an ancient bilaterian pathway supporting metabolic adaptation through STBD1 structural evolution

The selective autophagy of glycogen (glycophagy) has recently emerged as being crucial to glucose homeostasis in vertebrates, yet its origins remain elusive. Here, we provide evidence that starch-binding domain-containing protein 1 (STBD1), the key glycophagy receptor in vertebrates, is functionally conserved in the Pacific oyster, revealing its conserved position within ancient autophagy networks. We show that STBD1 in oysters - as seen in other invertebrate groups - possesses an N-terminal carbohydrate binding module family 20 (CBM20) domain, representing the ancestral state for this protein, while a shuffling of CBM20 to the C- terminus occurred during early chordate evolution. Structural modelling and functional studies reveal that the N-terminal CBM20 organization of STBD1 enhances glycogen binding. Functional experiments demonstrate that an STBD1-glycogen complex, anchored by GABARAPL2, facilitates an increased glycogen flux into autophagosomes for lysosomal degradation. We conclude that glycophagy is deeply conserved in bilaterians and that STBD1 structural evolution underlies potentially adaptive variation in metabolic strategies across distinct animal clades.

evolutionary biology↗
bioRxiv2024

NOLC1 Suppresses Immuno-chemotherapy by Inhibiting p53-mediated Ferroptosis in Gastric Cancer

Gastric cancer (GC) is one of the most malignant cancers, and cisplatin (Cis)-based chemotherapy remains the main clinical treatment for GC. However, Cis resistance often occurs, largely limiting its therapeutic efficacy in tumors. Therefore, a better understanding of the drug resistance mechanism could reveal new approaches for improving GC treatment efficacy. Here, we define the integrative role of nucleolar and coiled-body phosphoprotein 1 (NOLC1), a molecular chaperone that is significantly upregulated in GC tissues and Cis-resistant GC cells. Knocking down NOLC1 increased GC sensitivity to Cis by regulating ferroptosis. Mechanistically, NOLC1 binds to the p53 DNA binding domain (DBD), decreasing p53 nuclear accumulation stimulated by Cis and suppressing p53 transcriptional functions. Then, the p53-mediated ferroptosis is suppressed. Furthermore, the silence of NOLC1 promoted ferroptosis-induced immunogenic cell death (ICD) and reprogrammed the immunosuppressive tumor microenvironment, thereby increasing sensitivity to anti-programmed cell death-1 (PD-1) therapy plus Cis. The combination of anti-PD-1 plus Cis effectively inhibited GC growth without significant side effects. In summary, our findings reveal that targeting NOLC1 may be a novel therapeutic strategy for GC and may increase the efficacy of chemotherapy combined with immune checkpoint inhibitor (ICI) therapy.

cancer biology↗
bioRxiv2024

Deficiency of Hv1 Proton Channel Enhances Macrophage Antigen Presentation and Anti-Tumor Responses in Glioma

In the tumor microenvironment of glioblastoma, myeloid cells act as a double-edged sword--they are a major cellular component modulating the immune response while presenting a potential therapeutic target. Our study highlights the voltage-gated proton channel Hv1, predominantly expressed in myeloid cells, as a crucial regulator of their physiological functions. We discovered a strong correlation between increased Hv1 expression and poor prognosis in glioblastoma patients. Depleting Hv1 significantly extended survival in a mouse model of glioma. Employing multiple novel transgenic mouse lines, we demonstrated that Hv1 is upregulated in response to tumor presence, with glioma-associated macrophages as the principal contributors. Specifically, we identified that Hv1 in infiltrating macrophages as the major driver of survival phenotype differences. Through a combination of in vivo two-photon imaging, spectral flow cytometry, and spatial transcriptomic sequencing, we found that Hv1 depletion leads to reduced macrophage infiltration and enhanced antigen presentation, ultimately fostering a stronger adaptive immune response. These findings establish the Hv1 channel as a crucial new immune regulator within the brain tumor milieu, offering a promising target for reprogramming macrophage function to combat glioblastoma.

neuroscience↗
bioRxiv2024

SAMPL-seq reveals micron-scale spatial hubs in the human gut microbiome

The local arrangement of microbes can profoundly impact community assembly, function, and stability. To date, little is known about the spatial organization of the human gut microbiome. Here, we describe a high-throughput and streamlined method, dubbed SAMPL-seq, that samples microbial composition of micron-scale sub-communities with split-and-pool barcoding to capture spatial colocalization in a complex consortium. SAMPL-seq analysis of the gut microbiome of healthy humans identified bacterial taxa pairs that consistently co-occurred both over time and across multiple individuals. These colocalized microbes organize into spatially distinct groups or "spatial hubs" dominated by Bacteroideceae, Ruminococceae, and Lachnospiraceae families. From a dietary perturbation using inulin, we observed reversible spatial rearrangement of the gut microbiome, where specific taxa form new local partnerships. Spatial metagenomics using SAMPL-seq can unlock new insights to improve the study of microbial communities. One Sentence SummaryHigh throughput micron-scale subcommunity sampling and sequencing identifies distinct spatial associations of gut bacteria within and across individuals.

systems biology↗
bioRxiv2024

Perinatal Reduction of Genetically Aberrant Neurons from Human Cerebral Cortex

Human neurons are postmitotic and long-lived, requiring precise genomic regulation to maintain function over a lifetime. Normal neuronal function is highly dependent on gene dosage, with copy number variants (CNVs) and heterozygous point mutations associated with a host of neurodevelopmental and neuropsychiatric conditions [1-5]. Here, we investigated the landscape of somatic CNVs arising in fetal human brains, and how they change over development, to understand the processes that generate normal neuronal genomes. We sequenced 2,158 single neurons from human postmortem brain using two distinct single cell whole genome sequencing (scWGS) approaches. Tn5-transposase based (TbA) scWGS of 1,327 neurons from 16 individuals ranging in age from gestational week 14 to 90 years old resulted in 8,765 CNVs. Primary template amplification (PTA) was used to assess for CNV from 831 neurons from 12 individuals. Up to 46% of neurons in prenatal cortex showed aberrant genomes, characterized by widespread CNVs of multiple chromosomes, but this proportion reduces by 4-5 fold after birth across the two scWGS approaches (p=7.5x10-6, Fisher meta-analysis). We identified micronuclei in the developing cortex in situ, reflecting chromosomal material missegregated during neurodevelopment [6-8]. Neurons with widespread CNVs were eliminated in the perinatal period, while neurons with smaller CNV burden slowly declined during postnatal aging. CNVs in surviving neurons were depleted for genes that are dosage-sensitive or involved in neurodevelopmental disorders (p<0.05), suggesting selective elimination of neurons with CNVs involving these critical genes. We surveyed 44,861 nuclei with 10X Genomics scATAC/RNAseq and determined that neurons with high CNV burdens also showed abnormal expression of synaptic gene sets, suggesting that abnormal synaptic gene regulation contributes to neuronal elimination. Elimination of defective neuronal genomes during synaptogenesis may represent a critical process of genome quality control and a vulnerable target of factors that contribute to neurodevelopmental disease.

neuroscience↗
bioRxiv2024

SIRT4 enhances the cytotoxicity of NK cells toward hepatic stellate cells and reverses liver fibrosis via AMPKα/P-p53/NKG2DL pathway

Natural killer (NK) cells exhibit antifibrotic properties in liver fibrosis (LF) by suppressing activated hepatic stellate cells (HSCs). SIRT4, a mitochondrial regulatory protein, plays a crucial role as a link between energy metabolism and cell viability. However, the role of SIRT4 in the cytotoxicity of NK cells toward HSCs remains unexplored. In this study, we found that SIRT4 was markedly downregulated in both mouse models and patients with LF. The loss of SIRT4 reduced the cytotoxicity of NK cells against activated HSCs, while its overexpression enhanced this cytotoxicity. Mechanistically, SIRT4 activates AMPK to promote p53 phosphorylation and facilitates its nuclear translocation, which induces the transcription of ULBP1 and ULBP2, members of the NK group 2D Legend (NKG2DL) family of molecules. Finally, overexpression of SIRT4 activated mouse hepatic NK cells and reversed LF by constructing adeno-associated viruses (AAV) that specifically target HSCs. Thus, SIRT4 is essential for the cytotoxicity of NK cells toward HSCs, and AAV8-pGAFP-SIRT4 may serve as a therapeutic approach for managing LF. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=199 SRC="FIGDIR/small/615768v1_ufig1.gif" ALT="Figure 1"> View larger version (55K): org.highwire.dtl.DTLVardef@1e6e987org.highwire.dtl.DTLVardef@edcc2dorg.highwire.dtl.DTLVardef@1f7c2bcorg.highwire.dtl.DTLVardef@111d81d_HPS_FORMAT_FIGEXP M_FIG C_FIG

cell biology↗
bioRxiv2024

Different signaling interpretations by PKC eta and theta control T cell function and exhaustion

Chronic antigen signaling drives CD8+ T cell exhaustion (TEX) in cancer and chronic infection. However, how the kinase cascades downstream of the T cell receptor drive exhaustion is not understood. We found that continuous agonism of protein kinase C (PKC) causes degradation of PKC theta, but not PKC eta, and induces terminal TEX cells. During chronic infection, PKC theta is necessary to maintain the progenitor exhausted (TEX-PROG) cells, and thus the antigen-specific T cell response, while agonism of PKC eta promotes terminal exhaustion (TEX-TERM) in vitro and in vivo. The cascades downstream of these kinases are distinct, with PKC theta promoting activity of canonical PKC targets in the MAPK and CDK families, while eta promotes activity of other targets, including casein kinase I G2 (CK1G2). Expression of an engineered, degradation-resistant PKC theta, or deletion of the gene encoding CK1G2, improves T cell function and tumor control. Our illustration of multiple therapeutic avenues arising from targeting PKC highlights its centrality in TEX differentiation and its clinical potential in cancer immunotherapy. HighlightsO_LIPKC theta sustains T cell function while PKC eta promotes terminal exhaustion C_LIO_LIPKC theta and eta drive distinct phospho-cascades to oppose each others differentiation instructions C_LIO_LIAn engineered, degradation-resistant PKC theta improves T cell responses in chronic infection and cancer C_LIO_LIAblation of kinase CK1G2 downstream of PKC eta improves anti-tumor T cell responses C_LI

immunology↗
bioRxiv2024

Expanding the CRISPR toolbox by engineering Cas12a orthologs of metagenomic discovery

Cas12a (Cpf1) is a CRISPR-associated nuclease with broad utility in genome editing and molecular diagnostic applications. However, the widespread adoption of CRISPR-Cas12a nucleases and their variants has been hindered by the requirement for a specific protospacer adjacent motif (PAM), relatively low CRISPR RNA (crRNA) activity and the inability to multiplex nucleic acid detection alone. To overcome these limitations, we employed a comprehensive framework combined with AlphaFold2 to de novo mine 1,261 previously unexploited Cas12a orthologs from the global microbiome. Following experimental analysis, we identified the most promising 21 Cas12a nuclease orthologs and designated them "Genie scissor 12" (Gs12). Our analysis uncovered two exceptional variants among these newly identified orthologs: Gs12-10, a first natural PAM-less Cas12a ortholog, which can recognize 52 distinct PAM types, representing a significant 1.8-fold expansion in recognition range compared to the relative LbCas12a PAM; and Gs12-7MAX, an engineered variant of Gs12-7 that exhibited 1.27-fold higher editing efficiency than enAsCas12a-HF. Furthermore, we harnessed Gs12-1, Gs12-4, Gs12-9, and Gs12-18, along with their corresponding engineered crRNAs, to develop a powerful four-channel multiplexed CRISPR-based nucleic acid detection system. The discovery of diverse functions in Cas12a offers a deeper understanding of the CRISPR/Cas12a family. Also, it holds great promise for expanding its applications and uncovering the untapped potential of other CRISPR/Cas systems. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=189 SRC="FIGDIR/small/615316v1_ufig1.gif" ALT="Figure 1"> View larger version (57K): org.highwire.dtl.DTLVardef@bee75forg.highwire.dtl.DTLVardef@1480f61org.highwire.dtl.DTLVardef@1e9cbc9org.highwire.dtl.DTLVardef@4211b2_HPS_FORMAT_FIGEXP M_FIG C_FIG

synthetic biology↗
bioRxiv2024

Constructing a Mouse Brain Atlas of Dendritic Microenvironments Helps Discover Hidden Associations Between Anatomical Layout, Projection Targets and Transcriptomic Profiles of Neurons

Digital brain atlases have become essential anatomical references for understanding the spatial and functional organization of brains. For mice, typical resources include the Allen Reference Atlas, the Allen Common Coordinate Framework (CCF), and their variants, like CCFv3. However, previous whole-brain atlases were constructed based on limited neuronal features, such as cell body (soma) density or average maps from collections of registered brain images, without considering the spatial organization of neuronal arbors. This study introduces a microenvironment representation that incorporates the morphological features of neighboring neurons to better quantify brain modularity. We generated a large dataset containing dendrites from 101,136 neurons across 111 mouse brains, covering 91% of non-ventricular, non-fiber-tract CCF regions, and constructed a multidimensional microenvironment feature map of the whole brain. Our findings reveal that the spatial organization of these microenvironments outperforms the CCFv3 and a state-of-the-art spatial transcriptomic cell atlas by providing complementary subregions within established regions, nearly doubling the total number of brain regions compared to CCFv3. In this way, our atlas enables the identification of previously unobserved neuron groupings or "subtypes". Our results also demonstrate that this microenvironment atlas enhances local spatial homogeneity while maintaining spatial differentiation within established CCF brain regions. For example, we found that the microenvironments of hippocampal neurons are correlated with axonal projection targets and improve the specificity of projection mapping, which implies the potential characterization of long-range axonal projections of mammalian neurons based on only local dendritic organization. The sub-parcellation of the caudoputamen (CP) aligns well with previous studies on projections, connectivity, and transcriptomics, revealing diverse input and output wiring patterns among CP subregions.

neuroscience↗
bioRxiv2024

Aging-associated vacuolation of multi-ciliated cells in the distal mouse oviduct reflects unique cell identity and luminal microenvironment

The female reproductive organs present with the earliest aging characteristics, such as a decline in fertility and estrous cyclicity. While age-related changes in the ovary are well-documented, it is unclear if any age-associated changes occur in the other female reproductive organs, such as the oviduct/fallopian tube. The recent recognition of the distal end of the fallopian tube as the tissue of origin of high-grade serous tubal ovarian carcinomas (HGSCs), and that its patient demographic is strongly biased to postmenopausal women, motivated us to investigate age-associated changes in this organ. At the distal end of aged oviducts in mice, we found vacuolated multi-ciliated cells (MCCs) with a severely apically displaced and deformed nucleus. This phenotype was unique to the distal oviduct epithelium -infundibulum (INF) and ampulla (AMP). Ovariectomy did not affect the timeline of MCC vacuolation, suggesting little involvement of ovulation and hormonal regulation. MCC vacuolation was induced in hypoxia or hydroxyurea treatments in in vitro organotypic culture of all oviduct regions, not limited to the INF/AMP epithelium. This suggests high oxygen demand in MCCs, compared to other cell types, and a uniquely stressed INF/AMP epithelial microenvironment in vivo. We found that the blood circulation of INF/AMP depended on the ovarian artery, different from the rest of the oviduct epithelium and its circulation declined along with ovarian activities. We conclude that a decline in local blood circulation and distinct cellular identity of the INF/AMP epithelium caused age-associated MCC vacuolation, reflecting its mild, chronically stressed microenvironment.

physiology↗