Characterisation of a primary ciliary dyskinesia model generated from BMI1-transduced basal epithelial cells.

In this issue of Stem Cell Reports, Park et al. (2019) describe real-time in vivo visual monitoring of keratin-14+, Confetti-labeled limbal epithelial stem cells and their progeny as they contribute to central corneal wound-healing. The authors show that corneal wounds initially heal by “basal epithelial cell migration” into the wound-bed.

Basal airway epithelial cells (BCs) are multipotent stem cells key for epithelium homeostasis and repair. Despite their importance, little is known about BC phenotype in chronic infection in primary ciliary dyskinesia (PCD). Wild type (WT) and PCD basal cells were collected by nasal scrapings. PCD donors had bi-allelic mutations in DNAH11, DNAH5 and CCDC39. BCs were cultured ex vivo with either PBS or LPS (20 µg/mL). Post stimulation, cells were treated with either AZT (1 µg/mL or 10 µg/mL) or vehicle control. Cell proliferation was imaged for 72 hours. Supernatants were assayed for cytokines by 42-Luminex array at baseline and end. Proliferation was analyzed by linear mixed effects model, and cytokines by generalized maximum entropy estimation with p-values adjusted to control the false discovery rate at 5% across all cytokines. Treatment with AZT (1 µg/mL) significantly promoted cell proliferation in PCD BCs (p=0.02), while LPS-stimulation inhibited it (p<0.01). At baseline, there was significantly higher mean production of pro-inflammatory cytokines by CCDC39 than by WT, DNAH11 and DNAH5 BCs (p<0.05). LPS promoted significant increase in mean production of pro-inflammatory cytokines in PCD BCs (p<0.05). AZT treatment of LPS-stimulated PCD BCs reduced means of pro-inflammatory cytokines (p<0.05). We characterize inherent differences of PCD BCs in proliferation under control and pro-inflammatory conditions. In agreement with reported disease severity, we find significant differences in responses of CCDC39 BCs in comparison to WT, DNAH11 and DNAH5. Further, we find a significant effect of low dose AZT, providing rationale for AZT maintenance therapy in PCD.

Nasal Nitric Oxide Is an Important Test in the Diagnostic Pathway for Primary Ciliary Dyskinesia

The human heart is strikingly asymmetrical along the left-right body axis. If one begins with the position of the heart in the left chest, continues through asymmetrical venous drainage into the atria and asymmetrical orientation of the 2 anatomically and functionally distinct ventricles, and finally proceeds through the highly asymmetrical coil of semilunar valves and great vessels, the structure and function of the human heart are precisely aligned to the left-right axis. When cardiac asymmetries either fail to develop or align incorrectly relative to each other or relative to other organs, a plethora of congenital heart disease results. This group of heart diseases is called heterotaxy syndrome and represents both a difficult clinical challenge and a fascinating window into the biology underlying one of the most fundamental embryological processes, namely, the mechanism by which an organism establishes the 3 body axes. Positioning of organs along the left-right axis can be divided into 3 broad classes: situs solitus, in which all organs are positioned normally; situs inversus, in which there is mirror image reversal of all organs; and heterotaxy, in which there is any positioning of organs along the left-right axis differing from situs solitus and situs inversus (Figure). Pure situs inversus is found in 1 of 8500 in the general population and is usually not associated with intracardiac defects. In contrast, heterotaxy has a high degree of association with intracardiac defects. It has a reported incidence of 1 of 10 000 and is associated with at least 3% of cases of congenital heart disease.1 The anatomic spectrum of organ laterality. A, Situs solitus. B, Right atrial isomerism. The liver is midline, there are 2 eparterial bronchi, the position of the stomach and cardiac apex is indeterminate, and there is asplenia. C, Left atrial isomerism. The liver is midline, there …

Background: Primary ciliary dyskinesia (PCD) is a severe inherited disorder characterised by chronic respiratory disease, male infertility, and, in ∼50% of affected individuals, a left-right asymmetry defect called situs inversus....

Primary ciliary dyskinesia (PCD) is a genetic disorder associated with recurrent and chronic respiratory infections due to functional defects of motile cilia. In this study, we aimed to elucidate inflammatory and proliferative responses in PCD respiratory epithelium and evaluate the effect of Azithromycin (AZT) on these responses. Airway basal cells (BCs) were isolated from nasal samples of Wild-type (WT) epitope of healthy donors and PCD donors with bi-allelic mutations in DNAH5, DNAH11 and CCDC39. Cells were expanded in vitro and stimulated with either Lipopolysaccharide (LPS) or vehicle control. Post stimulation, cells were treated with either Azithromycin (AZT) or vehicle control. Cell proliferation was imaged in real-time. Separately, BCs from the same donors were expanded and grown at an air–liquid interface (ALI) to generate a multi-ciliated epithelium (MCE). Once fully mature, cells were stimulated with LPS, AZT, LPS + AZT or vehicle control. Inflammatory profiling was performed on collected media by cytokine Luminex assay. At baseline, there was a significantly higher mean production of pro-inflammatory cytokines by CCDC39 BCs and MCEs when compared to WT, DNAH11 and DNAH5 cells. AZT inhibited production of cytokines induced by LPS in PCD cells. Differences in cell proliferation were noted in PCD and this was also corrected with AZT treatment.

Whole-Exome Capture and Sequencing Identifies HEATR2 Mutation as a Cause of Primary Ciliary Dyskinesia

Super-Resolution Microscopy and FIB-SEM Imaging Reveal Parental Centriole-Derived, Hybrid Cilium in Mammalian Multiciliated Cells.

Carcinoma cells in PIN are situated above a layer of basal epithelial cells, which shield the tumor cells from stimulation by factors from the prostate stroma. During progression to invasive carcinoma, the basal cell layer becomes disrupted and tumor cells adhere to the basement membrane. The close proximity of basal epithelial cells to tumor cells in the early stages of prostate oncogenesis raises the possibility that basal epithelial cells participate in tumor cell invasion. Here, we investigated the migration-promoting activity of secreted factors from basal epithelial cells on BPH-1 cells, which we used as an in vitro model of preinvasive prostate cancer cells. We showed that the conditioned medium of basal epithelial cells (PEC-CM) contains adhesion proteins and chemotactic factors that stimulate adhesion, planar polarization, migration, and phosphorylation of Akt and that LY294002 and Wortmannin partially inhibit PEC-CM-triggered migration. We identified laminin-5 as a major migration-stimulating protein for BPH-1 cells in PEC-CM. Laminin-5 induced migration is completely inhibited by LY294002 or Wortmannin. In addition, antibody-depletion of laminin-5 from PEC-CM significantly diminishes the migration of BPH-1 cells. These results demonstrated, that laminin-5 is secreted by basal prostate epithelial cells in vivo and in vitro and stimulates migration of BPH-1 cells through a PI3-kinase dependent mechanism. Altogether, the possibility that basal epithelial cells assist in the invasion of in situ carcinoma cells is supported by the results from our in vitro system.

BACKGROUND The prostatic epithelium consists principally of basal epithelial cells, luminal epithelial cells, and neuroendocrine cells. Several studies support the concept that among basal cells, a subpopulation of stem cells resides which is capable of giving rise to other stem cells, basal epithelial cells, and also luminal epithelial cells and neuroendocrine cells. Other investigators suggest that luminal epithelial cells can also regenerate prostatic epithelium. Availability of pure populations of basal and luminal epithelial cells will aid in studies on defining the cellular pathways of differentiation during normal and pathological conditions. This study was designed to isolate and characterize pure populations of basal and luminal epithelial cells from adult rat ventral prostates. METHODS Sequential enzymatic digestion and differential plating permitted the separation of glandular epithelial cells from stromal cells. The glandular epithelial cells were subjected to the STAPUT technique. RESULTS Two types of cell populations, a large single-cell population and a small single-cell population, were obtained and characterized as basal and luminal epithelial cells by immunostaining for cytokeratin 5 and cytokeratin 8, respectively. CONCLUSIONS Our results indicate that purified populations of prostatic basal and luminal epithelial cells can be isolated by the STAPUT technique. Prostate 41:173–180, 1999. © 1999 Wiley-Liss, Inc.

The prostatic epithelium consists principally of basal epithelial cells, luminal epithelial cells, and neuroendocrine cells. Several studies support the concept that among basal cells, a subpopulation of stem cells resides which is capable of giving rise to other stem cells, basal epithelial cells, and also luminal epithelial cells and neuroendocrine cells. Other investigators suggest that luminal epithelial cells can also regenerate prostatic epithelium. Availability of pure populations of basal and luminal epithelial cells will aid in studies on defining the cellular pathways of differentiation during normal and pathological conditions. This study was designed to isolate and characterize pure populations of basal and luminal epithelial cells from adult rat ventral prostates. Sequential enzymatic digestion and differential plating permitted the separation of glandular epithelial cells from stromal cells. The glandular epithelial cells were subjected to the STAPUT technique. Two types of cell populations, a large single-cell population and a small single-cell population, were obtained and characterized as basal and luminal epithelial cells by immunostaining for cytokeratin 5 and cytokeratin 8, respectively. Our results indicate that purified populations of prostatic basal and luminal epithelial cells can be isolated by the STAPUT technique.

A sperm viability test using SYBR-14/propidium iodide flow cytometry as a tool for rapid screening of primary ciliary dyskinesia patients and for choosing sperm sources for intracytoplasmic sperm injection

Phosphotyrosine Antibodies Preferentially React with Basal Epithelial Cells in the Dog Prostate

Whole-exome sequencing identified novel DNAH5 homozygous variants in two consanguineous families with primary ciliary dyskinesia.

p63 Coordinates Anogenital Modeling and Epithelial Cell Differentiation in the Developing Female Urogenital Tract