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Cellular Atlas of Laryngeal and Voca...

Lunga, Tadeas.

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Cellular Atlas of Laryngeal and Vocal Fold Embryogenesis, Maturation and Aging.

Record Type:	Electronic resources : Monograph/item
Title/Author:	Cellular Atlas of Laryngeal and Vocal Fold Embryogenesis, Maturation and Aging./
Author:	Lunga, Tadeas.
Published:	Ann Arbor : ProQuest Dissertations & Theses, : 2024,
Description:	150 p.
Notes:	Source: Dissertations Abstracts International, Volume: 85-12, Section: A.
Contained By:	Dissertations Abstracts International85-12A.
Subject:	Developmental biology. -
Online resource:	https://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=31330283
ISBN:	9798382817903

Cellular Atlas of Laryngeal and Vocal Fold Embryogenesis, Maturation and Aging.
Lunga, Tadeas.

Cellular Atlas of Laryngeal and Vocal Fold Embryogenesis, Maturation and Aging. - Ann Arbor : ProQuest Dissertations & Theses, 2024 - 150 p.

Source: Dissertations Abstracts International, Volume: 85-12, Section: A.

Thesis (Ph.D.)--The University of Wisconsin - Madison, 2024.

IntroductionEmbryogenesis, maturation, and aging of the larynx and vocal folds (VF) are intricate processes crucial for voice production. Structural variations in VF mucosa across different age groups, from newborns to the elderly, predispose individuals to age-associated voice disorders. Treatment options for these disorders remain limited due to gaps in understanding the cellular and genetic processes underlying these structural changes. To address these gaps, our study aimed to identify and comprehend the cellular heterogeneity of VF across the lifespan using single-cell RNA sequencing. In addition, we aimed to identify differentially expressed genes across lifespan and developmental trajectories and relationships between different cell populations.MethodsWe used the Bl6 mouse strain, harvesting larynges at various embryonic (E) and postnatal (P) stages. Tissues were processed into single-cell suspensions, and cDNA libraries were prepared and sequenced. Data underwent quality control, normalization, integration, and clustering with annotations based on differentially expressed genes (DEG). In addition, we performed pseudotime trajectory analysis for epithelial and mesenchymal cell populations. We also performed DEG analysis across timepoints for major epithelial, mesenchymal, and immune cell populations.ResultsClustering identified 23 major cell populations, including epithelial, mesenchymal, endothelial, immune, and neuronal populations. Epithelial clusters included basement membrane-producing cells (BMPC), proliferating basal epithelial cells (PBEC), nonproliferating basal epithelial cells (NPBEC), suprabasal epithelial cells (SBEC), ciliated epithelial cells (CC), and secretory epithelial cells (SEC). Mesenchymal clusters included early proliferating mesenchymal progenitors (EPMP), stromal cells, fibroblasts (FB), chondroblasts, chondrocytes, smooth muscle cells, and skeletal muscle cells. Immune cell clusters comprised macrophages (MG), dendritic cells, neutrophils, and lymphocytes. Endothelial cells and neuronal clusters were also identified. Cell complexity increased from embryonic stages to adulthood, peaking between adolescence and adulthood before declining. Prenatal stages showed predominance of mesenchymal populations. Postnatal stages showed an increase in epithelial populations. In later postnatal stages there was an influx of immune cells specifically neutrophiles and pro-inflammatory macrophages. The DEG analysis across time-points revealed functional maturation of epithelial cells starting 4W of age in PBEC, NPBEC, and SBEC. FB showed collagen production maturation between E18.5 to 4W. Trajectory analysis revealed EPMP gave rise to chondroblasts, fibroblasts, and skeletal muscle cells. The PBEC was a precursor to major differentiated cell populations such as SBEC, SEC, and CC. Lastly, we revealed multiple subtypes for major cell populations such as BMPC, PBEC, NPBEC, SBEC, SEC, EPMP, FB, and MG.ConclusionOur single-cell atlas reveals the cellular heterogeneity of VF and laryngeal tissue throughout development. Major cell populations were established early, with mesenchymal populations predominantly in embryonic stages and epithelial cell populations in postnatal development. Epithelial cells mature at 4W. FBs mature earlier between E18.5 to 4W. The cell atlas of laryngeal and VF development across lifespan serves as a benchmark for designing novel engineered-based strategies for VF tissue restoration and/or replacements, differentiation trajectories, cross species tissue comparison and translational research focused on age-, gene-, and population-specific treatment options.

ISBN: 9798382817903Subjects--Topical Terms:

592588
Developmental biology.
Subjects--Index Terms:

Vocal folds

Cellular Atlas of Laryngeal and Vocal Fold Embryogenesis, Maturation and Aging.
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245 1 0 $a Cellular Atlas of Laryngeal and Vocal Fold Embryogenesis, Maturation and Aging.
260 1 $a Ann Arbor : $b ProQuest Dissertations & Theses, $c 2024
300 $a 150 p.
500 $a Source: Dissertations Abstracts International, Volume: 85-12, Section: A.
500 $a Advisor: Thibeault, Susan L.
502 $a Thesis (Ph.D.)--The University of Wisconsin - Madison, 2024.
520 $a IntroductionEmbryogenesis, maturation, and aging of the larynx and vocal folds (VF) are intricate processes crucial for voice production. Structural variations in VF mucosa across different age groups, from newborns to the elderly, predispose individuals to age-associated voice disorders. Treatment options for these disorders remain limited due to gaps in understanding the cellular and genetic processes underlying these structural changes. To address these gaps, our study aimed to identify and comprehend the cellular heterogeneity of VF across the lifespan using single-cell RNA sequencing. In addition, we aimed to identify differentially expressed genes across lifespan and developmental trajectories and relationships between different cell populations.MethodsWe used the Bl6 mouse strain, harvesting larynges at various embryonic (E) and postnatal (P) stages. Tissues were processed into single-cell suspensions, and cDNA libraries were prepared and sequenced. Data underwent quality control, normalization, integration, and clustering with annotations based on differentially expressed genes (DEG). In addition, we performed pseudotime trajectory analysis for epithelial and mesenchymal cell populations. We also performed DEG analysis across timepoints for major epithelial, mesenchymal, and immune cell populations.ResultsClustering identified 23 major cell populations, including epithelial, mesenchymal, endothelial, immune, and neuronal populations. Epithelial clusters included basement membrane-producing cells (BMPC), proliferating basal epithelial cells (PBEC), nonproliferating basal epithelial cells (NPBEC), suprabasal epithelial cells (SBEC), ciliated epithelial cells (CC), and secretory epithelial cells (SEC). Mesenchymal clusters included early proliferating mesenchymal progenitors (EPMP), stromal cells, fibroblasts (FB), chondroblasts, chondrocytes, smooth muscle cells, and skeletal muscle cells. Immune cell clusters comprised macrophages (MG), dendritic cells, neutrophils, and lymphocytes. Endothelial cells and neuronal clusters were also identified. Cell complexity increased from embryonic stages to adulthood, peaking between adolescence and adulthood before declining. Prenatal stages showed predominance of mesenchymal populations. Postnatal stages showed an increase in epithelial populations. In later postnatal stages there was an influx of immune cells specifically neutrophiles and pro-inflammatory macrophages. The DEG analysis across time-points revealed functional maturation of epithelial cells starting 4W of age in PBEC, NPBEC, and SBEC. FB showed collagen production maturation between E18.5 to 4W. Trajectory analysis revealed EPMP gave rise to chondroblasts, fibroblasts, and skeletal muscle cells. The PBEC was a precursor to major differentiated cell populations such as SBEC, SEC, and CC. Lastly, we revealed multiple subtypes for major cell populations such as BMPC, PBEC, NPBEC, SBEC, SEC, EPMP, FB, and MG.ConclusionOur single-cell atlas reveals the cellular heterogeneity of VF and laryngeal tissue throughout development. Major cell populations were established early, with mesenchymal populations predominantly in embryonic stages and epithelial cell populations in postnatal development. Epithelial cells mature at 4W. FBs mature earlier between E18.5 to 4W. The cell atlas of laryngeal and VF development across lifespan serves as a benchmark for designing novel engineered-based strategies for VF tissue restoration and/or replacements, differentiation trajectories, cross species tissue comparison and translational research focused on age-, gene-, and population-specific treatment options.
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653 $a Voice
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793 $a English
856 4 0 $u https://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=31330283