Tu1734 Inhibition of MRP4 Enhances the Wound Repair Process:

Wound repair is a key feature distinguishing living from nonliving matter. Single cells are increasingly recognized to be capable of healing wounds. The lack of reproducible, high-throughput wounding methods has hindered single-cell wound repair studies. This work describes a microfluidic guillotine for bisecting single Stentor coeruleus cells in a continuous-flow manner. Stentor is used as a model due to its robust repair capacity and the ability to perform gene knockdown in a high-throughput manner. Local cutting dynamics reveals two regimes under which cells are bisected, one at low viscous stress where cells are cut with small membrane ruptures and high viability and one at high viscous stress where cells are cut with extended membrane ruptures and decreased viability. A cutting throughput up to 64 cells per minute—more than 200 times faster than current methods—is achieved. The method allows the generation of more than 100 cells in a synchronized stage of their repair process. This capacity, combined with high-throughput gene knockdown in Stentor, enables time-course mechanistic studies impossible with current wounding methods.

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Cell wound repair in Drosophila occurs through three distinct phases of membrane and cytoskeletal remodeling

The Journal of Cell Biology ◽

10.1083/jcb.201011018 ◽

2011 ◽

Vol 193 (3) ◽

pp. 455-464 ◽

Cited By ~ 78

Author(s):

Maria Teresa Abreu-Blanco ◽

Jeffrey M. Verboon ◽

Susan M. Parkhurst

Keyword(s):

Cell Death ◽

Plasma Membrane ◽

Single Cell ◽

Wound Repair ◽

Single Cells ◽

Repair Process ◽

Wound Edge ◽

Cytoskeletal Remodeling ◽

Tissue Integrity ◽

E Cadherin

When single cells or tissues are injured, the wound must be repaired quickly in order to prevent cell death, loss of tissue integrity, and invasion by microorganisms. We describe Drosophila as a genetically tractable model to dissect the mechanisms of single-cell wound repair. By analyzing the expression and the effects of perturbations of actin, myosin, microtubules, E-cadherin, and the plasma membrane, we define three distinct phases in the repair process—expansion, contraction, and closure—and identify specific components required during each phase. Specifically, plasma membrane mobilization and assembly of a contractile actomyosin ring are required for this process. In addition, E-cadherin accumulates at the wound edge, and wound expansion is excessive in E-cadherin mutants, suggesting a role for E-cadherin in anchoring the actomyosin ring to the plasma membrane. Our results show that single-cell wound repair requires specific spatial and temporal cytoskeleton responses with distinct components and mechanisms required at different stages of the process.

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Overcoming stromal barriers to immuno-oncological responses via fibroblast activation protein-targeted therapy

Immunotherapy ◽

10.2217/imt-2020-0066 ◽

2021 ◽

Vol 13 (2) ◽

pp. 155-175

Author(s):

W Nathaniel Brennen ◽

Daniel L J Thorek ◽

Wen Jiang ◽

Timothy E Krueger ◽

Lizamma Antony ◽

...

Keyword(s):

Prostate Cancer ◽

Targeted Therapy ◽

Immune Suppression ◽

Feedback Loop ◽

Wound Repair ◽

Repair Process ◽

Fibroblast Activation Protein ◽

Primary Data ◽

Positive Feedback Loop ◽

Fibroblast Activation

The tumor microenvironment contributes to disease progression through multiple mechanisms, including immune suppression mediated in part by fibroblast activation protein (FAP)-expressing cells. Herein, a review of FAP biology is presented, supplemented with primary data. This includes FAP expression in prostate cancer and activation of latent reservoirs of TGF-β and VEGF to produce a positive feedback loop. This collectively suggests a normal wound repair process subverted during cancer pathophysiology. There has been immense interest in targeting FAP for diagnostic, monitoring and therapeutic purposes. Until recently, this development has outpaced an understanding of the biology; impeding optimal translation into the clinic. A summary of these applications is provided with an emphasis on eliminating tumor-infiltrating FAP-positive cells to overcome stromal barriers to immuno-oncological responses.

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Histological response to injury in the horseshoe crab, Limulus polyphemus

Canadian Journal of Zoology ◽

10.1139/z77-150 ◽

1977 ◽

Vol 55 (7) ◽

pp. 1158-1165 ◽

Cited By ~ 18

Author(s):

Charles R. Bursey

Keyword(s):

Epithelial Cells ◽

Wound Repair ◽

Horseshoe Crab ◽

Thick Layer ◽

Repair Process ◽

Limulus Polyphemus ◽

Tissue Structure ◽

Histological Response ◽

Wound Site

Observations are presented on the wound repair process of the horseshoe crab, Limulus polyphemus. A cut, 1 × 40 × 3 mm, was made through the dorsal abdominal carapace and observations of the tissue at the wound site were made at 6, 24, 48, 96 h and 5, 10, 15, 30, 60, 90, 120, and 180 days. The wound was immediately plugged by a hemolymph coagulum and there was heavy infiltration of hemocytes into the area. Infiltrating hemocytes undergo a series of changes. The outermost cells hyalinize and form a thick layer between the cut ends of the exoskeleton. Pigmented epithelial cells migrate into the scar and produce a layer of cuticle under the cut exoskeleton. During the remaining period of observation, the cuticular scab enlarged and the cellular mass that filled the wound channel was lost through a series of changes that eventually restored the original tissue structure.

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ECRG4 regulates neutrophil recruitment and CD44 expression during the inflammatory response to injury

Science Advances ◽

10.1126/sciadv.aay0518 ◽

2020 ◽

Vol 6 (11) ◽

pp. eaay0518 ◽

Cited By ~ 1

Author(s):

Robert A. Dorschner ◽

Jisook Lee ◽

Olga Cohen ◽

Todd Costantini ◽

Andrew Baird ◽

...

Keyword(s):

Wound Repair ◽

Repair Process ◽

Neutrophil Recruitment ◽

Related Gene ◽

Cd44 Expression ◽

Cancer Related Gene ◽

Inflammation Resolution ◽

Inflammatory Response To Injury ◽

Gene 4

The complex molecular microenvironment of the wound bed regulates the duration and degree of inflammation in the wound repair process, while its dysregulation leads to impaired healing. Understanding factors controlling this response provides therapeutic targets for inflammatory disease. Esophageal cancer–related gene 4 (ECRG4) is a candidate chemokine that is highly expressed on leukocytes. We used ECRG4 knockout (KO) mice to establish that the absence of ECRG4 leads to defective neutrophil recruitment with a delay in wound healing. An in vitro human promyelocyte model identified an ECRG4-mediated suppression of the hyaluronic acid receptor, CD44, a key receptor mediating inflammation resolution. In ECRG4 KO mouse leukocytes, there was an increase in CD44 expression, consistent with a model in which ECRG4 negatively regulates CD44 levels. Therefore, we propose a previously unidentified mechanism in which ECRG4 regulates early neutrophil recruitment and subsequent CD44-mediated resolution of inflammation.

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Senescence of IPF Lung Fibroblasts Disrupt Alveolar Epithelial Cell Proliferation and Promote Migration in Wound Healing

Pharmaceutics ◽

10.3390/pharmaceutics12040389 ◽

2020 ◽

Vol 12 (4) ◽

pp. 389 ◽

Cited By ~ 3

Author(s):

Kaj E. C. Blokland ◽

David W. Waters ◽

Michael Schuliga ◽

Jane Read ◽

Simon D. Pouwels ◽

...

Keyword(s):

Cell Proliferation ◽

Epithelial Cell ◽

Wound Repair ◽

Alveolar Epithelial Cell ◽

Lung Parenchyma ◽

Repair Process ◽

Lung Fibroblasts ◽

Epithelial Cell Proliferation ◽

Alveolar Epithelial ◽

M Phase

Idiopathic pulmonary fibrosis (IPF) is a progressive lung disease marked by excessive accumulation of lung fibroblasts (LFs) and collagen in the lung parenchyma. The mechanisms that underlie IPF pathophysiology are thought to reflect repeated alveolar epithelial injury leading to an aberrant wound repair response. Recent work has shown that IPF-LFs display increased characteristics of senescence including growth arrest and a senescence-associated secretory phenotype (SASP) suggesting that senescent LFs contribute to dysfunctional wound repair process. Here, we investigated the influence of senescent LFs on alveolar epithelial cell repair responses in a co-culture system. Alveolar epithelial cell proliferation was attenuated when in co-culture with cells or conditioned media from, senescence-induced control LFs or IPF-LFs. Cell-cycle analyses showed that a larger number of epithelial cells were arrested in G2/M phase when co-cultured with IPF-LFs, than in monoculture. Paradoxically, the presence of LFs resulted in increased A549 migration after mechanical injury. Our data suggest that senescent LFs may contribute to aberrant re-epithelialization by inhibiting proliferation in IPF.

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Nanofibers for improving the wound repair process: the combination of a grafted chitosan and an antioxidant agent

Polymer Chemistry ◽

10.1039/c7py00038c ◽

2017 ◽

Vol 8 (10) ◽

pp. 1664-1671 ◽

Cited By ~ 32

Author(s):

Lan Mei ◽

Rangrang Fan ◽

Xiaoling Li ◽

Yuelong Wang ◽

Bo Han ◽

...

Keyword(s):

Wound Healing ◽

Wound Repair ◽

Repair Process ◽

Complex Process ◽

Antioxidant Agent

Wound healing, a complex process involving several important biomolecules and pathways, requires efficient dressings to enhance the therapy effects.

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Toll-Like Receptor Agonists Modulate Wound Regeneration in Airway Epithelial Cells

International Journal of Molecular Sciences ◽

10.3390/ijms19082456 ◽

2018 ◽

Vol 19 (8) ◽

pp. 2456 ◽

Cited By ~ 4

Author(s):

Anna Lewandowska-Polak ◽

Małgorzata Brauncajs ◽

Marzanna Jarzębska ◽

Małgorzata Pawełczyk ◽

Marcin Kurowski ◽

...

Keyword(s):

Epithelial Cells ◽

Mrna Expression ◽

Airway Epithelium ◽

Wound Repair ◽

Repair Process ◽

Airway Epithelial Cells ◽

Poly I:C ◽

Toll Like Receptors ◽

Vegf Mrna ◽

Microbial Products

Background: Impaired regeneration of airway epithelium may lead to persistence of inflammation and remodelling. Regeneration of injured epithelium is a complex phenomenon and the role of toll-like receptors (TLRs) in the stimulation of respiratory virus products in this process has not been established. Objective: This study was undertaken to test the hypothesis that the wound repair process in airway epithelium is modulated by microbial products via toll-like receptors. Methods: Injured and not-injured bronchial epithelial cells (ECs) (BEAS-2B line) were incubated with the TLR agonists poly(I:C), lipopolisacharide (LPS), allergen Der p1, and supernatants from virus-infected epithelial cells, either alone or in combination with TLR inhibitors. Regeneration and immune response in injured and not-injured cells were studied. Results: Addition of either poly(I:C) or LPS to ECs induced a marked inhibition of wound repair. Supernatants from RV1b-infected cells also decreased regeneration. Preincubation of injured and not-injured ECs with TLR inhibitors decreased LPS and poly(I:C)-induced repair inhibition. TGF-β and RANTES mRNA expression was higher in injured ECs and IFN-α, IFN-β, IL-8, and VEGF mRNA expression was lower in damaged epithelium as compared to not-injured. Stimulation with poly(I:C) increased IFN-α and IFN-β mRNA expression in injured cells, and LPS stimulation decreased interferons mRNA expression both in not-injured and injured ECs. Conclusion: Regeneration of the airway epithelium is modulated by microbial products via toll-like receptors.

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Cnidarian Immunity and the Repertoire of Defense Mechanisms in Anthozoans

Biology ◽

10.3390/biology9090283 ◽

2020 ◽

Vol 9 (9) ◽

pp. 283

Author(s):

Maria Giovanna Parisi ◽

Daniela Parrinello ◽

Loredana Stabili ◽

Matteo Cammarata

Keyword(s):

Intracellular Signaling ◽

Defense Mechanisms ◽

Wound Repair ◽

Gene Families ◽

Repair Process ◽

Immune Defense ◽

Current State ◽

Self Recognition ◽

Essential Components ◽

Broad Array

Anthozoa is the most specious class of the phylum Cnidaria that is phylogenetically basal within the Metazoa. It is an interesting group for studying the evolution of mutualisms and immunity, for despite their morphological simplicity, Anthozoans are unexpectedly immunologically complex, with large genomes and gene families similar to those of the Bilateria. Evidence indicates that the Anthozoan innate immune system is not only involved in the disruption of harmful microorganisms, but is also crucial in structuring tissue-associated microbial communities that are essential components of the cnidarian holobiont and useful to the animal’s health for several functions including metabolism, immune defense, development, and behavior. Here, we report on the current state of the art of Anthozoan immunity. Like other invertebrates, Anthozoans possess immune mechanisms based on self/non-self-recognition. Although lacking adaptive immunity, they use a diverse repertoire of immune receptor signaling pathways (PRRs) to recognize a broad array of conserved microorganism-associated molecular patterns (MAMP). The intracellular signaling cascades lead to gene transcription up to endpoints of release of molecules that kill the pathogens, defend the self by maintaining homeostasis, and modulate the wound repair process. The cells play a fundamental role in immunity, as they display phagocytic activities and secrete mucus, which acts as a physicochemical barrier preventing or slowing down the proliferation of potential invaders. Finally, we describe the current state of knowledge of some immune effectors in Anthozoan species, including the potential role of toxins and the inflammatory response in the Mediterranean Anthozoan Anemonia viridis following injection of various foreign particles differing in type and dimensions, including pathogenetic bacteria.

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