Actin polymerization inhibition by targeting ARPC2 affects intestinal stem cell homeostasis

Ruzhen Zhang; Sheng Chen; Zhifan Yang; Ning Zhang; Kenan Guo; Keyi Lv; Zimo Zhou; Meijiao Gao; Xiancheng Hu; Yongping Su; Jianming He; Fengchao Wang

doi:10.1093/burnst/tkad038

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Actin polymerization inhibition by targeting ARPC2 affects intestinal stem cell homeostasis

更新时间：2024-02-04

- Actin polymerization inhibition by targeting ARPC2 affects intestinal stem cell homeostasis
- Actin polymerization inhibition by targeting ARPC2 affects intestinal stem cell homeostasis
- Burns & Trauma 2023年11卷第1期
- 作者机构：
  
  1. Third Military Medical University
  2. Chongqing Normal University
  3. Hebei University of Chinese Medicine, Key Laboratory of Integrated Chinese and Western Medicine for Gastroenterology Research,Hebei
- 作者简介：
- 基金信息：
  
  This research was supported by the National Natural Science Foundation of China (81872556);Chongqing Academician Program (Basic Research and Frontier Exploration) cstc2018jcyj-yszxX0004.
- DOI：10.1093/burnst/tkad038
  中图分类号：
- 纸质出版日期：2023，
  
  网络出版日期：2023-10-16，
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Ruzhen Zhang, Sheng Chen, Zhifan Yang, 等. Actin polymerization inhibition by targeting ARPC2 affects intestinal stem cell homeostasis[J]. Burns & Trauma, 2023,11(1).

Ruzhen Zhang, Sheng Chen, Zhifan Yang, Ning Zhang, Kenan Guo, Keyi Lv, Zimo Zhou, Meijiao Gao, Xiancheng Hu, Yongping Su, Jianming He, Fengchao Wang, Actin polymerization inhibition by targeting ARPC2 affects intestinal stem cell homeostasis, Burns & Trauma, Volume 11, 2023, tkad038, https://doi.org/10.1093/burnst/tkad038
Ruzhen Zhang, Sheng Chen, Zhifan Yang, 等. Actin polymerization inhibition by targeting ARPC2 affects intestinal stem cell homeostasis[J]. Burns & Trauma, 2023,11(1). DOI： 10.1093/burnst/tkad038.

Ruzhen Zhang, Sheng Chen, Zhifan Yang, Ning Zhang, Kenan Guo, Keyi Lv, Zimo Zhou, Meijiao Gao, Xiancheng Hu, Yongping Su, Jianming He, Fengchao Wang, Actin polymerization inhibition by targeting ARPC2 affects intestinal stem cell homeostasis, Burns & Trauma, Volume 11, 2023, tkad038, https://doi.org/10.1093/burnst/tkad038 DOI：

摘要

Abstract

BackgroundThe rapid turnover of the intestinal epithelium is driven by the proliferation and differentiation of intestinal stem cells (ISCs). The dynamics of the F-actin cytoskeleton are critical for maintaining intercellular force and the signal transduction network. However

it remains unclear how direct interference with actin polymerization impacts ISC homeostasis. This study aims to reveal the regulatory effects of the F-actin cytoskeleton on the homeostasis of intestinal epithelium

as well as the potential risks of benproperine (BPP) as an anti-tumor drug.MethodsPhalloidin fluorescence staining was utilized to test F-actin polymerization. Flow cytometry and IHC staining were employed to discriminate different types of intestinal epithelial cells. Cell proliferation was assessed through bromo-deoxyuridine (BrdU) and 5-ethynyl-2′-deoxyuridine (EdU) incorporation assays. The proliferation and differentiation of intestinal stem cells were replicated in vitro through organoid culture. Epithelial migration was evaluated through BrdU pulse labeling and chasing in mice.ResultsThe F-actin content was observed to significantly increase as crypt cells migrated into the villus region. Additionally

actin polymerization in secretory cells

especially in Paneth cells (PCs)

was much higher than that in neighboring ISCs. Treatment with the newly identified actin-related protein 2/3 complex subunit 2 (ARPC2) inhibitor BPP led to a dose-dependent increase or inhibition of intestinal organoid growth in vitro and crypt cell proliferation in vivo. Compared with the vehicle group

BPP treatment decreased the expression of Lgr5 ISC feature genes in vivo and in organoid culture. Meanwhile

PC differentiation derived from ISCs and progenitors was decreased by inhibition of F-actin polymerization. Mechanistically

BPP-induced actin polymerization inhibition may activate the Yes1-associated transcriptional regulator pathway

which affects ISC proliferation and differentiation. Accordingly

BPP treatment affected intestinal epithelial cell migration in a dose-dependent manner.ConclusionOur findings indicate that the regulation of cytoskeleton reorganization can affect ISC homeostasis. In addition

inhibiting ARPC2 with the Food and Drug Administration-approved drug BPP represents a novel approach to influencing the turnover of intestinal epithelial cells.

关键词

Keywords

F-actinBenproperineARPC2Intestinal stem cellYAP

references

1.Clevers H. The intestinal crypt, a prototype stem cell compartment. Cell. 2013;154:274–84.

2.Sato T, van Es JH, Snippert HJ, Stange DE, Vries RG, van den Born M, et al. Paneth cells constitute the niche for Lgr5 stem cells in intestinal crypts. Nature. 2011;469:415–8.

3.Beumer J, Clevers H. Cell fate specification and differentiation in the adult mammalian intestine. Nat Rev Mol Cell Biol. 2021;22:39–53.

4.Moujaber O, Stochaj U. The cytoskeleton as regulator of cell Signaling pathways. Trends Biochem Sci. 2020;45:96–107.

5.Pocaterra A, Santinon G, Romani P, Brian I, Dimitracopoulos A, Ghisleni A, et al. F-actin dynamics regulates mammalian organ growth and cell fate maintenance. J Hepatol. 2019;71:130–42.

6.Aloisio FM, Barber DL. Arp2/3 complex activity is necessary for mouse ESC differentiation, times formative pluripotency, and enables lineage specification. Stem Cell Reports. 2022;17:1318–33.

7.Chen L, Hu H, Qiu W, Shi K, Kassem M. Actin depolymerization enhances adipogenic differentiation in human stromal stem cells. Stem Cell Res. 2018;29:76–83.

8.Sen B, Uzer G, Samsonraj RM, Xie Z, McGrath C, Styner M, et al. Intranuclear actin structure modulates mesenchymal stem cell differentiation. Stem Cells. 2017;35:1624–35.

9.Hogrebe NJ, Augsornworawat P, Maxwell KG, Velazco-Cruz L, Millman JR. Targeting the cytoskeleton to direct pancreatic differentiation of human pluripotent stem cells. Nat Biotechnol. 2020;38:460-+.

10.Sun B, Qu R, Fan T, Yang Y, Jiang X, Khan AU, et al. Actin polymerization state regulates osteogenic differentiation in human adipose-derived stem cells. Cell Mol Biol Lett. 2021;26:15.

11.Izdebska M, Zielinska W, Halas-Wisniewska M, Grzanka A. Involvement of actin and actin-binding proteins in carcinogenesis. Cell. 2020;9102245.

12.Heng YW, Koh CG. Actin cytoskeleton dynamics and the cell division cycle. Int J Biochem Cell Biol. 2010;42:1622–33.

13.Molinie N, Gautreau A. THE Arp2/3 REGULATORY SYSTEM AND ITS DEREGULATION IN CANCER. Physiol Rev. 2018;98:215–38.

14.Chanez-Paredes S, Montoya-Garcia A, Schnoor M. Cellular and pathophysiological consequences of Arp2/3 complex inhibition: role of inhibitory proteins and pharmacological compounds. Cell Mol Life Sci. 2019;76:3349–61.

15.Hetrick B, Han MS, Helgeson LA, Nolen BJ. Small molecules CK-666 and CK-869 inhibit actin-related protein 2/3 complex by blocking an activating conformational change. Chem Biol. 2013;20:701–12.

16.Yoon YJ, Han YM, Choi J, Lee YJ, Yun J, Lee SK, et al. Benproperine, an ARPC2 inhibitor, suppresses cancer cell migration and tumor metastasis. Biochem Pharmacol. 2019;163:46–59.

17.Ren J, Niu Z, Li X, Yang J, Gao M, Li X, et al. A novel morphometry system automatically assessing the growth and regeneration of intestinal organoids. Biochem Biophys Res Commun. 2018;506:1052–8.

18.Wang F, Scoville D, He XC, Mahe MM, Box A, Perry JM, et al. Isolation and characterization of intestinal stem cells based on surface marker combinations and colony-formation assay. Gastroenterology. 2013;145:e1–21.

19.Faust JJ, Millis BA, Tyska MJ. Profilin-mediated actin allocation regulates the growth of epithelial microvilli. Curr Biol. 2019;29:3457–3465 e3.

20.Holzinger A. Jasplakinolide: an actin-specific reagent that promotes actin polymerization. Methods Mol Biol. 2009;586:71–87.

21.Barker N, van Es JH, Kuipers J, Kujala P, van den Born M, Cozijnsen M, et al. Identification of stem cells in small intestine and colon by marker gene Lgr5. Nature. 2007;449:1003–7.

22.Snippert HJ, van der Flier LG, Sato T, van Es JH, van den Born M, Kroon-Veenboer C, et al. Intestinal crypt homeostasis results from neutral competition between symmetrically dividing Lgr5 stem cells. Cell. 2010;143:134–44.

23.Gregorieff A, Liu Y, Inanlou MR, Khomchuk Y, Wrana JL. Yap-dependent reprogramming of Lgr5(+) stem cells drives intestinal regeneration and cancer. Nature. 2015;526:715–8.

24.Krndija D, El Marjou F, Guirao B, Richon S, Leroy O, Bellaiche Y, et al. Active cell migration is critical for steady-state epithelial turnover in the gut. Science. 2019;365:705-+.

25.Chanez-Paredes S, Montoya-Garcia A, Castro-Ochoa KF, Garcia-Cordero J, Cedillo-Barron L, Shibayama M, et al. The Arp2/3 inhibitory protein Arpin is required for intestinal epithelial barrier integrity. Front Cell Dev Biol. 2021;9:625719.

26.Wang F, Wang F, Zou Z, Liu D, Wang J, Su Y. Active deformation of apoptotic intestinal epithelial cells with adhesion-restricted polarity contributes to apoptotic clearance. Lab Investig. 2011;91:462–71.

27.De Belly H, Paluch EK, Chalut KJ. Interplay between mechanics and signalling in regulating cell fate. Nat Rev Mol Cell Biol. 2022;23:465–80.

28.Bottcher A, Buttner M, Tritschler S, Sterr M, Aliluev A, Oppenlander L, et al. Non-canonical Wnt/PCP signalling regulates intestinal stem cell lineage priming towards enteroendocrine and Paneth cell fates. Nat Cell Biol. 2021;23:23–31.

29.Gracz AD, Williamson IA, Roche KC, Johnston MJ, Wang F, Wang Y, et al. A high-throughput platform for stem cell niche co-cultures and downstream gene expression analysis. Nat Cell Biol. 2015;17:340–9.

30.Yu S, Tong K, Zhao Y, Balasubramanian I, Yap GS, Ferraris RP, et al. Paneth cell multipotency induced by notch activation following injury. Cell Stem Cell. 2018;23:e5.

31.Zhang W, Liao Y, Lou J, Zhuang M, Yan H, Li Q, et al. CircRNA_Maml2 promotes the proliferation and migration of intestinal epithelial cells after severe burns by regulating the miR-93-3p/FZD7/Wnt/beta-catenin pathway. Burns. Trauma. 2022;10:tkac009.

32.Azzolin L, Panciera T, Soligo S, Enzo E, Bicciato S, Dupont S, et al. YAP/TAZ incorporation in the beta-catenin destruction complex orchestrates the Wnt response. Cell. 2014;158:157–70.

33.Damkham N, Issaragrisil S, Lorthongpanich C. Role of YAP as a Mechanosensing molecule in stem cells and stem cell-derived hematopoietic cells. Int J Mol Sci. 2022;2314634.

34.Barry ER, Morikawa T, Butler BL, Shrestha K, de la Rosa R, Yan KS, et al. Restriction of intestinal stem cell expansion and the regenerative response by YAP. Nature 2013;493:106–10.

35.Liu M, Zhang Z, Sampson L, Zhou X, Nalapareddy K, Feng YX, et al. RHOA GTPase controls YAP-mediated EREG Signaling in small intestinal stem cell maintenance. Stem Cell Reports. 2017;9:1961–75.

36.Halder G, Dupont S, Piccolo S. Transduction of mechanical and cytoskeletal cues by YAP and TAZ. Nat Rev Mol Cell Biol. 2012;13:591–600.

37.Dupont S, Morsut L, Aragona M, Enzo E, Giulitti S, Cordenonsi M, et al. Role of YAP/TAZ in mechanotransduction. Nature. 2011;474:179–83.

38.Zhang Z, Zhang F, Davis AK, Xin M, Walz G, Tian W, et al. CDC42 controlled apical-basal polarity regulates intestinal stem cell to transit amplifying cell fate transition via YAP-EGF-mTOR signaling. Cell Rep. 2022;38:110009.

39.Watson JR, Owen D, Mott HR. Cdc42 in actin dynamics: an ordered pathway governed by complex equilibria and directional effector handover. Small GTPases. 2017;8:237–44.

40.Rauhala HE, Teppo S, Niemela S, Kallioniemi A. Silencing of the ARP2/3 complex disturbs pancreatic cancer cell migration. Anticancer Res. 2013;33:45–52.

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