Limbal BCAM expression identifies a proliferative progenitor population capable of holoclone formation and corneal differentiation

Sasamoto, Yuzuru; Lee, Catherine A. A.; Wilson, B. J.; Buerger, Florian; Martin, Gabrielle; Kiritoshi, Shoko; Tran, Johnathan; González, Gabriel; Hildebrandt, Friedhelm; Jo, Vickie Y.; Lian, Christine G.; Murphy, Gëorge F.; Ksander, Bruce R.; Frank, Markus H.; Frank, Natasha Y.

doi:10.1016/j.celrep.2022.111166

Cited by 9 publications

(6 citation statements)

References 46 publications

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“…Moreover, we found that human BCs from CRSwNP express higher levels of p63-dependent genes. CHIP-seq data demonstrate that p63 binds to a region in the BCAM promoter (90), and a recent study identified that overexpression of Np63 in 293T cells increases luciferase activity in a reporter containing the BCAM promoter region (91). Thus, the reproducibility with which BCAM expression identifies airway stem cells across species and conditions may reflect its regulation by p63.…”

Section: Discussionmentioning

confidence: 99%

Type 2 Inflammation Drives an Airway Basal Stem Cell Program Through Insulin Receptor Substrate Signaling

Wang

Hallen

Lee

et al. 2022

Preprint

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Background: Chronic rhinosinusitis with nasal polyposis (CRSwNP) is a type 2 (T2) inflammatory disease associated with an increased number of airway basal epithelial cells (BCs). Recent studies have identified transcriptionally distinct BCs, but functional data are lacking and the molecular pathways that support or inhibit human BC proliferation and differentiation are largely unknown. Objective: To determine the role of T2 cytokines in regulating airway BCs. Methods: Single cell and bulk RNA-sequencing of sinus and lung airway epithelial cells was analyzed. Human sinus BCs were stimulated with IL-4 and IL-13 in the presence and absence of IL4R inhibitors. Confocal analysis of human sinus tissue and murine airway was performed. Murine BC subsets were sorted for RNA sequencing and functional assays. Fate labeling was performed in a murine model of tracheal injury and repair. Results: Here we find two subsets of BCs in human and murine respiratory mucosa distinguished by the expression of BC adhesion molecule (BCAM). BCAM expression identifies airway stem cells among P63+KRT5+NGFR+ BCs. In the sinonasal mucosa, BCAMhi BCs expressing TSLP, IL33, CCL26, and the canonical BC transcription factor TP63 are increased in patients with CRSwNP. In cultured BCs, IL-4/13 increases expression of BCAM and TP63 through an Insulin Receptor Substrate (IRS)-dependent signaling pathway that is increased in CRSwNP. Conclusions: These findings establish BCAM as a marker of airway stem cells among the BC pool and demonstrate that airway epithelial remodeling in T2 inflammation extends beyond goblet cell metaplasia to the support of a BC stem state poised to perpetuate inflammation.

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Section: Discussionmentioning

confidence: 99%

Type 2 Inflammation Drives an Airway Basal Stem Cell Program Through Insulin Receptor Substrate Signaling

Wang

Hallen

Lee

et al. 2022

Preprint

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“…Formalin-fixed paraffin-embedded sections were subjected to deparaffinization and antigen retrieval steps before immunostaining as described [12]. The tissue sections were permeabilized with Triton-X (MilliporeSigma) containing buffer and blocked with 5-10% normal donkey or goat serum (Jackson ImmunoResearch Laboratories, West Grove, PA) or 1% bovine serum albumin (BSA) (MilliporeSigma) containing buffer.…”

Section: Immunostainingmentioning

confidence: 99%

“…LSCs located at the basal epithelial layer in the outermost part of the cornea express ABCB5 and p63 [2][3][4][5][6][7][8][9][10]. Transit amplifying cells occupy the basal epithelial layer inside the limbal circle and are characterized by expression of ß-integrin and BCAM [11,12]. Terminally differentiated corneal epithelial cells are found in the apical layers of the central cornea, where they 2 of 16 express MUC16 [13].…”

Section: Introductionmentioning

confidence: 99%

Epigenetic Regulation of Corneal Epithelial Differentiation by TET2

Sasamoto

Lee

et al. 2023

IJMS

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Epigenetic DNA modification by 5-hydroxymethylcytosine (5hmC), generated by the Ten-eleven translocation (TET) dioxygenases, regulates diverse biological functions in many organ tissues, including the mammalian eye. For example, 5hmC has been shown to be involved in epigenetic regulation of retinal gene expression. However, a functional role of 5hmC in corneal differentiation has not been investigated to date. Here, we examined 5hmC and TET function in the human cornea. We found 5hmC highly expressed in MUC16-positive terminally differentiated cells that also co-expressed the 5hmC-generating enzyme TET2. TET2 knockdown (KD) in cultured corneal epithelial cells led to significant reductions of 5hmC peak distributions and resulted in transcriptional repression of molecular pathways involved in corneal differentiation, as evidenced by downregulation of MUC4, MUC16, and Keratin 12. Additionally, integrated TET2 KD RNA-seq and genome-wide Reduced Representation Hydroxymethylation Profiling revealed novel epigenetically regulated genes expressed by terminally differentiated cells, including KRT78, MYEOV, and MAL. In aggregate, our findings reveal a novel function of TET2 in the epigenetic regulation of corneal epithelial gene expression and identify novel TET2-controlled genes expressed in differentiated corneal epithelial cells. These results point to potential roles for TET2 induction strategies to enhance treatment of corneal diseases associated with abnormal epithelial maturation.

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“…Shalom‐Feuerstein et al 6 reported that p63‐deficient mice exhibited disrupted corneal epithelial cell (CEC) differentiation and stratification. Recently, several factors have been implicated in LSC expansion in vitro, including basal cell adhesion molecule, 7 tetraspanin 7, 8 and SRY‐box transcription factor 2, 9 however, the underlying regulatory mechanism of LSC proliferation remains largely unclear.…”

Section: Introductionmentioning

confidence: 99%

“…Various approaches have been used to explore the mechanism of epithelial cell proliferation, including transgenic mouse models, 6 lineage-tracking technology, 1,10 holoclone formation in cell culture, 11 and three-dimensional 5 or air-lifting 7 corneal-epithelial differentiation systems. It is worth noting that epithelial stem/progenitor cells exhibit diverse proliferation capacities and an overall proliferation decline during cell division in vitro.…”

mentioning

confidence: 99%

Single‐cell transcriptomics implicates the FEZ1–DKK1 axis in the regulation of corneal epithelial cell proliferation and senescence

Zhu

Wang

et al. 2023

Cell Proliferation

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Limbal stem/progenitor cells (LSC) represent the source of corneal epithelium renewal. LSC proliferation and differentiation are essential for corneal homeostasis, however, the regulatory mechanism remains largely unexplored. Here, we performed single‐cell RNA sequencing and discovered proliferation heterogeneity as well as spontaneously differentiated and senescent cell subgroups in multiply passaged primary LSC. Fasciculation and elongation protein zeta 1 (FEZ1) and Dickkopf‐1 (DKK1) were identified as two significant regulators of LSC proliferation and senescence. These two factors were mainly expressed in undifferentiated corneal epithelial cells (CECs). Knocking down the expression of either FEZ1 or DKK1 reduced cell division and caused cell cycle arrest. We observed that DKK1 acted as a downstream target of FEZ1 in LSC and that exogenous DKK1 protein partially prevented growth arrest and senescence upon FEZ1 suppression in vitro. In a mouse model of corneal injury, DKK1 also rescued the corneal epithelium after recovery was inhibited by FEZ1 suppression. Hence, the FEZ1–DKK1 axis was required for CEC proliferation and the juvenile state and can potentially be targeted as a therapeutic strategy for promoting recovery after corneal injury.

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Limbal BCAM expression identifies a proliferative progenitor population capable of holoclone formation and corneal differentiation

Cited by 9 publications

References 46 publications

Type 2 Inflammation Drives an Airway Basal Stem Cell Program Through Insulin Receptor Substrate Signaling

Type 2 Inflammation Drives an Airway Basal Stem Cell Program Through Insulin Receptor Substrate Signaling

Epigenetic Regulation of Corneal Epithelial Differentiation by TET2

Single‐cell transcriptomics implicates the FEZ1–DKK1 axis in the regulation of corneal epithelial cell proliferation and senescence

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