Single-cell atlas of healthy vocal folds and cellular function in the endothelial-to-mesenchymal transition

Danling Liu , Yunzhong Zhang , Luo Guo , Rui Fang , Jin Guo , Peifang Li , Tingting Qian , Wen Li , Liping Zhao , Xiaoning Luo , Siyi Zhang , Jun Shao , Shan Sun

Cell Proliferation ›› 2024, Vol. 57 ›› Issue (12) : e13723

PDF
Cell Proliferation ›› 2024, Vol. 57 ›› Issue (12) : e13723 DOI: 10.1111/cpr.13723
ORIGINAL ARTICLE

Single-cell atlas of healthy vocal folds and cellular function in the endothelial-to-mesenchymal transition

Author information +
History +
PDF

Abstract

The vocal fold is an architecturally complex organ comprising a heterogeneous mixture of various layers of individual epithelial and mesenchymal cell lineages. Here we performed single-cell RNA sequencing profiling of 5836 cells from the vocal folds of adult Sprague–Dawley rats. Combined with immunostaining, we generated a spatial and transcriptional map of the vocal fold cells and characterized the subpopulations of epithelial cells, mesenchymal cells, endothelial cells, and immune cells. We also identified a novel epithelial-to-mesenchymal transition-associated epithelial cell subset that was mainly found in the basal epithelial layers. We further confirmed that this subset acts as intermediate cells with similar genetic features to epithelial-to-mesenchymal transition in head and neck squamous cell carcinoma. Finally, we present the complex intracellular communication network involved homeostasis using CellChat analysis. These studies define the cellular and molecular framework of the biology and pathology of the VF mucosa and reveal the functional importance of developmental pathways in pathological states in cancer.

Cite this article

Download citation ▾
Danling Liu, Yunzhong Zhang, Luo Guo, Rui Fang, Jin Guo, Peifang Li, Tingting Qian, Wen Li, Liping Zhao, Xiaoning Luo, Siyi Zhang, Jun Shao, Shan Sun. Single-cell atlas of healthy vocal folds and cellular function in the endothelial-to-mesenchymal transition. Cell Proliferation, 2024, 57(12): e13723 DOI:10.1111/cpr.13723

登录浏览全文

4963

注册一个新账户 忘记密码

References

[1]

LungovaV, Thibeault SL. Mechanisms of larynx and vocal fold development and pathogenesis. Cell Mol Life Sci. 2020; 77(19):3781-3795.

[2]

LiL, Stiadle JM, LauHK, et al. Tissue engineering-based therapeutic strategies for vocal fold repair and regeneration. Biomaterials. 2016; 108:91-110.

[3]

KumaiY. Pathophysiology of fibrosis in the vocal fold: current research, future treatment strategies, and obstacles to restoring vocal fold pliability. Int J Mol Sci. 2019; 20(10):1-18.

[4]

SmithG. Structure of the normal rat larynx. Lab Anim. 1977; 11(4):223-228.

[5]

LiX, WangCY. From bulk, single-cell to spatial RNA sequencing. Int J Oral Sci. 2021; 13(1):36.

[6]

HedlundE, DengQ. Single-cell RNA sequencing: technical advancements and biological applications. Mol Aspects Med. 2018; 59:36-46.

[7]

BusslingerGA, Weusten BLA, BogteA, BegthelH, Brosens LAA, CleversH. Human gastrointestinal epithelia of the esophagus, stomach, and duodenum resolved at single-cell resolution. Cell Rep. 2021; 34(10):108819.

[8]

EsworthyRS, Doroshow JH, ChuFF. The beginning of GPX2 and 30 years later. Free Radic Biol Med. 2022; 188:419-433.

[9]

KaluckaJ, de Rooij L, GoveiaJ, et al. Single-cell transcriptome atlas of murine endothelial cells. Cell. 2020; 180(4):764-779 e720.

[10]

MuhlL, GenoveG, LeptidisS, et al. Single-cell analysis uncovers fibroblast heterogeneity and criteria for fibroblast and mural cell identification and discrimination. Nat Commun. 2020; 11(1):3953.

[11]

DengCC, HuYF, ZhuDH, et al. Single-cell RNA-seq reveals fibroblast heterogeneity and increased mesenchymal fibroblasts in human fibrotic skin diseases. Nat Commun. 2021; 12(1):3709.

[12]

MorrishE, RulandJ. Cytotoxic FCER1G(+) innate-like T cells: new potential for tumour immunotherapy. Signal Transduct Target Ther. 2022; 7(1):204.

[13]

GirblT, LennT, PerezL, et al. Distinct compartmentalization of the chemokines CXCL1 and CXCL2 and the atypical receptor ACKR1 determine discrete stages of neutrophil Diapedesis. Immunity. 2018; 49(6):1062-1076.e1066.

[14]

Le VercheV, Sunshine SS, HammersD, SweeneyHL, Paushkin S. Chapter 21 - Skeletal Muscle in Spinal Muscular Atrophy As an Opportunity for Therapeutic Intervention. In: CJ Sumner, S Paushkin, C-P Ko, eds. Spinal Muscular Atrophy. Academic Press; 2017:341-356.

[15]

SincennesMC, BrunCE, LinAYT, et al. Acetylation of PAX7 controls muscle stem cell self-renewal and differentiation potential in mice. Nat Commun. 2021; 12(1):3253.

[16]

LiW, Danilenko DM, BuntingS, et al. The serine protease marapsin is expressed in stratified squamous epithelia and is up-regulated in the hyperproliferative epidermis of psoriasis and regenerating wounds. J Biol Chem. 2009; 284(1):218-228.

[17]

BitonM, HaberAL, RogelN, et al. T helper cell cytokines modulate intestinal stem cell renewal and differentiation. Cell. 2018; 175(5):1307-1320.e1322.

[18]

DelgadoR, MuraCV, BacigalupoJ. Single Ca(2+)-activated Cl(–) channel currents recorded from toad olfactory cilia. BMC Neurosci. 2016; 17(1):17.

[19]

MuraCV, Delgado R, DelgadoMG, RestrepoD, Bacigalupo J. A CLCA regulatory protein present in the chemosensory cilia of olfactory sensory neurons induces a Ca(2+)-activated Cl(–) current when transfected into HEK293. BMC Neurosci. 2017; 18(1):61.

[20]

KathiriyaJJ, WangC, ZhouM, et al. Human alveolar type 2 epithelium transdifferentiates into metaplastic KRT5(+) basal cells. Nat Cell Biol. 2022; 24(1):10-23.

[21]

YinL, LiQ, MrdenovicS, et al. KRT13 promotes stemness and drives metastasis in breast cancer through a plakoglobin/c-Myc signaling pathway. Breast Cancer Res. 2022; 24(1):7.

[22]

WangN, LinKK, LuZ, et al. The LIM-only factor LMO4 regulates expression of the BMP7 gene through an HDAC2-dependent mechanism, and controls cell proliferation and apoptosis of mammary epithelial cells. Oncogene. 2007; 26(44):6431-6441.

[23]

FarrL, GhoshS, JiangN, et al. CD74 signaling links inflammation to intestinal epithelial cell regeneration and promotes mucosal healing. Cell Mol Gastroenterol Hepatol. 2020; 10(1):101-112.

[24]

HouY, ZhouY, JehiL, et al. Aging-related cell type-specific pathophysiologic immune responses that exacerbate disease severity in aged COVID-19 patients. Aging Cell. 2022; 21(2):e13544.

[25]

RawlinsEL, OkuboT, XueY, et al. The role of Scgb1a1+ Clara cells in the long-term maintenance and repair of lung airway, but not alveolar, epithelium. Cell Stem Cell. 2009; 4(6):525-534.

[26]

KimuraS, Yokoyama S, PilonAL, KurotaniR. Emerging role of an immunomodulatory protein secretoglobin 3A2 in human diseases. Pharmacol Ther. 2022; 236:108112.

[27]

ChenW, YangA, JiaJ, PopovYV, SchuppanD, You H. Lysyl oxidase (LOX) family members: rationale and their potential as therapeutic targets for liver fibrosis. Hepatology. 2020; 72(2):729-741.

[28]

MilewiczDM, Braverman AC, De BackerJ, et al. Nat Rev Dis Primers. Marfan Syndrome. 2021; 7(1):64.

[29]

NandaV, MianoJM. Leiomodin 1, a new serum response factor-dependent target gene expressed preferentially in differentiated smooth muscle cells. J Biol Chem. 2012; 287(4):2459-2467.

[30]

NandaV, WangT, PjanicM, et al. Functional regulatory mechanism of smooth muscle cell-restricted LMOD1 coronary artery disease locus. PLoS Genet. 2018; 14(11):e1007755.

[31]

LamouilleS, XuJ, DerynckR. Molecular mechanisms of epithelial-mesenchymal transition. Nat Rev Mol Cell Biol. 2014; 15(3):178-196.

[32]

PuramSV, TiroshI, ParikhAS, et al. Single-cell transcriptomic analysis of primary and metastatic tumor ecosystems in head and neck cancer. Cell. 2017; 171(7):1611-1624.e1624.

[33]

PastushenkoI, Blanpain C. EMT transition states during tumor progression and metastasis. Trends Cell Biol. 2019; 29(3):212-226.

[34]

LarruceaS, ButtaN, RodriguezRB, et al. Podocalyxin enhances the adherence of cells to platelets. Cell Mol Life Sci. 2007; 64(22):2965-2974.

[35]

LutterS, XieS, TatinF, Makinen T. Smooth muscle-endothelial cell communication activates Reelin signaling and regulates lymphatic vessel formation. J Cell Biol. 2012; 197(6):837-849.

[36]

YuH, TangD, WuH, et al. Integrated single-cell analyses decode the developmental landscape of the human fetal spine. iScience. 2022; 25(7):104679.

[37]

RoumeliotisS, Dounousi E, EleftheriadisT, LiakopoulosV. Association of the inactive circulating matrix gla protein with vitamin K intake, calcification, mortality, and cardiovascular disease: a review. Int J Mol Sci. 2019; 20(3):1-27.

[38]

Tsuji-TamuraK, Morino-Koga S, SuzukiS, OgawaM. The canonical smooth muscle cell marker TAGLN is present in endothelial cells and is involved in angiogenesis. J Cell Sci. 2021; 134(15):1-8.

[39]

JitprapaikulsanJ, Klein CJ, PittockSJ, et al. Phenotypic presentations of paraneoplastic neuropathies associated with MAP1B-IgG. J Neurol Neurosurg Psychiatry. 2020; 91(3):328-330.

[40]

XiaoX, YeohBS, Vijay-KumarM. Lipocalin 2: an emerging player in iron homeostasis and inflammation. Annu Rev Nutr. 2017; 37:103-130.

[41]

JacksonRM, Griesel BA, ShortKR, SparlingD, Freeman WM, OlsonAL. Weight loss results in increased expression of anti-inflammatory protein CRISPLD2 in mouse adipose tissue. Obesity (Silver Spring). 2019; 27(12):2025-2036.

[42]

LiF, ZhuW, GonzalezFJ. Potential role of CYP1B1 in the development and treatment of metabolic diseases. Pharmacol Ther. 2017; 178:18-30.

[43]

WuJ, ChuX, ChenC, Bellusci S. Role of fibroblast growth factor 10 in mesenchymal cell differentiation during lung development and disease. Front Genet. 2018; 9:545.

[44]

NyengP, BjerkeMA, NorgaardGA, Qu X, KobberupS, JensenJ. Fibroblast growth factor 10 represses premature cell differentiation during establishment of the intestinal progenitor niche. Dev Biol. 2011; 349(1):20-34.

[45]

AndoK, WangW, PengD, et al. Peri-arterial specification of vascular mural cells from naïve mesenchyme requires Notch signaling. Development. 2019; 146(2):1-13.

[46]

MazzoniJ, SmithJR, ShahriarS, Cutforth T, CejaB, AgalliuD. The Wnt inhibitor Apcdd1 coordinates vascular remodeling and barrier maturation of retinal blood vessels. Neuron. 2017; 96(5):1055-1069.e1056.

[47]

LaitmanBM, Charytonowicz D, ZhuAJ, et al. High-resolution profiling of human vocal fold cellular landscapes with single-nuclei RNA sequencing. Laryngoscope. 2024; 134(7):3193-3200.

[48]

DeprezM, Zaragosi LE, TruchiM, et al. A single-cell atlas of the human healthy airways. Am J Respir Crit Care Med. 2020; 202(12):1636-1645.

[49]

DongreA, Weinberg RA. New insights into the mechanisms of epithelial-mesenchymal transition and implications for cancer. Nat Rev Mol Cell Biol. 2019; 20(2):69-84.

[50]

SatoK, Chitose SI, SatoK, SatoF, OnoT, UmenoH. Glycolysis of tissue stem cells in the macula Flava of newborn vocal fold. J Voice. 2022.

[51]

YouD, GuoJ, ZhangY, et al. The heterogeneity of mammalian utricular cells over the course of development. Clin Transl Med. 2022; 12(10):e1052.

RIGHTS & PERMISSIONS

2024 The Author(s). Cell Proliferation published by Beijing Institute for Stem Cell and Regenerative Medicine and John Wiley & Sons Ltd.

AI Summary AI Mindmap
PDF

218

Accesses

0

Citation

Detail

Sections
Recommended

AI思维导图

/