GO:1903690 negative regulation of wound healing, spreading of epidermal cells: Mechanism, Genes, Functions and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:1903690 describes any process that stops, prevents, or reduces the frequency, rate, or extent of wound healing, spreading of epidermal cells.
• Epidermal cell spreading during wound healing is driven by actin cytoskeletal remodeling and focal adhesion turnover, processes controlled by Rho-family GTPases such as Rac1.
• Rac1 activity in epidermal stem cells is required for migration and re-epithelialization, and its modulation directly affects wound healing outcomes.
• Deleted in Liver Cancer 1 (DLC1) is a Rho GTPase-activating protein (GAP) that can be activated by epidermal growth factor (EGF) through focal adhesion kinase (FAK) and protein phosphatase 2A (PP2A), linking growth factor signaling to negative regulation of cell spreading.
• Dysregulation of epidermal wound healing processes is associated with chronic inflammatory skin diseases such as psoriasis, where DNA methylation patterns in CD4+ T-cells distinguish patients from healthy controls.
• CRISPR-based knockout, point mutation, knock-in, and overexpression models enable causal testing of candidate genes in the GO:1903690 pathway.
Description
The Gene Ontology term GO:1903690, negative regulation of wound healing, spreading of epidermal cells, defines any biological process that stops, prevents, or reduces the frequency, rate, or extent of epidermal cell spreading during wound healing. This term captures the braking mechanisms that ensure epidermal sheet migration is spatially and temporally controlled, preventing excessive or disorganized re-epithelialization. Epidermal spreading is a critical step in wound closure, requiring coordinated actin cytoskeletal dynamics, focal adhesion turnover, and cell-matrix interactions. The small GTPase Rac1 is a central regulator of epidermal stem cell migration during wound healing, and its modulation directly influences the rate of epidermal sheet expansion. Negative regulation of this process is equally important, as unrestrained epidermal migration can lead to pathological outcomes including chronic wounds and inflammatory skin diseases. The Rho GTPase-activating protein Deleted in Liver Cancer 1 (DLC1) provides a molecular link between growth factor signaling and suppression of cell spreading, as epidermal growth factor (EGF) activates DLC1 via focal adhesion kinase (FAK) and protein phosphatase 2A (PP2A). Understanding the negative regulators of epidermal wound healing is essential for developing therapies that promote timely wound closure while preventing aberrant epidermal proliferation. This article synthesizes the current knowledge of GO:1903690, its molecular players, disease relevance, and the CRISPR-based research methods used to study it.
negative regulation of wound healing, spreading of epidermal cells At A Glance
| GO ID | GO:1903690 |
|---|---|
| GO term | negative regulation of wound healing, spreading of epidermal cells |
| Ontology | biological_process |
| Synonym | down regulation of wound healing, spreading of epidermal cells; down-regulation of wound healing, spreading of epidermal cells; downregulation of wound healing, spreading of epidermal cells; inhibition of wound healing, spreading of epidermal cells |
| Major function | Suppression of epidermal cell spreading and migration during wound healing |
| Key regulators | Rac1, DLC1, FAK, PP2A, EGF signaling |
| Related process | Regulation of actin cytoskeleton organization, focal adhesion dynamics |
| Disease relevance | Psoriasis, chronic wounds, impaired re-epithelialization |
What Is GO:1903690?
GO:1903690 is a biological process term defined as any process that stops, prevents, or reduces the frequency, rate, or extent of wound healing, spreading of epidermal cells. In other words, it encompasses the molecular and cellular mechanisms that put the brakes on the migration and spreading of epidermal cells during the wound healing response. This includes signaling events that inhibit actin-driven protrusion, promote focal adhesion disassembly, or otherwise restrain the locomotory behavior of epidermal keratinocytes and stem cells.
Why Is negative regulation of wound healing, spreading of epidermal cells Important in Cell Biology?
GO:1903690 is important because the negative regulation of epidermal cell spreading is essential for proper wound healing; without it, epidermal migration can become excessive or disorganized, leading to chronic wounds, scarring, or inflammatory skin diseases such as psoriasis. Understanding these inhibitory mechanisms provides targets for therapeutic intervention to promote wound closure in chronic wounds or to restrain pathological epidermal activation in psoriasis. Moreover, the molecular players involved, such as Rac1 and DLC1, are conserved regulators of cell migration with broad relevance to cancer metastasis and tissue repair.
• Controls the spatial and temporal extent of epidermal sheet migration during wound closure.
• Prevents excessive or disorganized re-epithelialization that can lead to scarring.
• Rac1 modulation in epidermal stem cells directly affects wound healing rate.
• DLC1 activation by EGF via FAK and PP2A provides a mechanism for growth factor-dependent inhibition of cell spreading.
• Dysregulation is linked to chronic inflammatory skin diseases such as psoriasis.
• Provides potential therapeutic targets for chronic wounds and psoriasis.
• Shares molecular machinery with cancer cell migration and metastasis.
• Enables CRISPR-based functional genomics to identify novel negative regulators.
What Happens During negative regulation of wound healing, spreading of epidermal cells?
Initiation of epidermal cell spreading
In simple terms: Epidermal cells start to move and spread to cover a wound.
Upon wounding, epidermal stem cells and keratinocytes at the wound edge become activated and begin to migrate into the wound bed. This process requires dynamic reorganization of the actin cytoskeleton and formation of new focal adhesions. Rac1, a Rho-family GTPase, is a key driver of this migratory response in epidermal stem cells.
Negative regulation by Rho GTPase-activating proteins
In simple terms: Proteins called GAPs can shut down the signals that tell cells to move.
Negative regulation of epidermal cell spreading can be achieved by GTPase-activating proteins (GAPs) that inactivate Rac1 and related GTPases. Deleted in Liver Cancer 1 (DLC1) is a Rho GTPase-activating protein that can suppress cell spreading when activated. This provides a molecular brake on epidermal migration.
Growth factor signaling to DLC1
In simple terms: Growth factors can turn on the brake signal through a chain of proteins.
Epidermal growth factor (EGF) activates DLC1 via focal adhesion kinase (FAK) and protein phosphatase 2A (PP2A). This signaling cascade links extracellular growth factor cues to the negative regulation of cell spreading, ensuring that epidermal migration is tightly controlled by the wound microenvironment.
Focal adhesion turnover and cytoskeletal remodeling
In simple terms: The cell's attachment points and internal skeleton are constantly rebuilt to control movement.
Negative regulation of epidermal cell spreading involves changes in focal adhesion dynamics and actin cytoskeletal remodeling. DLC1 activation by EGF/FAK/PP2A signaling can lead to reduced focal adhesion turnover and decreased protrusive activity, thereby limiting cell spreading. Rac1 activity is also modulated to balance migration and stasis.
Termination of epidermal migration
In simple terms: Once the wound is covered, the cells stop moving.
As wound healing progresses, negative regulatory mechanisms ensure that epidermal cell spreading is terminated once the wound is closed. This prevents overgrowth and maintains tissue homeostasis. Dysregulation of these stopping signals can contribute to chronic wounds or inflammatory skin diseases.
Key Genes Involved in GO:1903690 negative regulation of wound healing, spreading of epidermal cells
The following genes and proteins have been implicated in the negative regulation of wound healing, spreading of epidermal cells (GO:1903690) based on published literature.
| Gene | Major Role | Research Relevance |
|---|---|---|
| RAC1 | Rho-family GTPase driving epidermal stem cell migration; its modulation affects wound healing rate | Target for studying positive and negative regulation of epidermal spreading |
| DLC1 | Rho GTPase-activating protein (GAP) activated by EGF/FAK/PP2A; suppresses cell spreading | Key negative regulator; potential tumor suppressor |
| FAK (PTK2) | Focal adhesion kinase; mediates EGF-induced DLC1 activation | Links growth factor signaling to negative regulation of spreading |
| PPP2CA | Protein phosphatase 2A catalytic subunit; required for EGF-induced DLC1 activation | Phosphatase involved in negative regulation |
| EGF | Epidermal growth factor; triggers signaling cascade that activates DLC1 | Extracellular cue for negative regulation |
| EGFR | Epidermal growth factor receptor; upstream of FAK/PP2A/DLC1 pathway | Receptor tyrosine kinase mediating EGF effects |
| CD4 | T-cell marker; DNA methylation patterns in CD4+ T-cells distinguish psoriasis patients | Immune cell link to epidermal wound healing dysregulation |
| RAC1B | Alternatively spliced Rac1 isoform; may modulate Rac1 activity | Potential modifier of epidermal migration |
| ARHGAP22 | Rho GTPase-activating protein; may regulate Rac1 and cell spreading | Candidate negative regulator |
| ARHGAP24 | Rho GTPase-activating protein; involved in cytoskeletal regulation | Candidate negative regulator |
| SRC | Non-receptor tyrosine kinase; can regulate FAK and focal adhesions | Potential modulator of DLC1 activation |
| PTEN | Phosphatase and tensin homolog; regulates PIP3 and cell migration | Indirect regulator of epidermal spreading |
| PIK3CA | PI3-kinase catalytic subunit; downstream of growth factor signaling | Modulates Rac1 activation |
| CDC42 | Rho-family GTPase; regulates filopodia and cell migration | Related to Rac1 in epidermal spreading |
| RHOA | Rho-family GTPase; regulates stress fibers and focal adhesions | Balances Rac1-driven migration |
| VCL | Vinculin; focal adhesion protein | Marker of focal adhesion dynamics |
| ACTB | Beta-actin; cytoskeletal component | Readout of cytoskeletal remodeling |
| ITGB1 | Integrin beta 1; mediates cell-matrix adhesion | Required for epidermal cell spreading |
How Is negative regulation of wound healing, spreading of epidermal cells Regulated?
The negative regulation of wound healing, spreading of epidermal cells is controlled by a signaling axis involving epidermal growth factor (EGF), focal adhesion kinase (FAK), protein phosphatase 2A (PP2A), and the Rho GTPase-activating protein DLC1. EGF stimulation leads to FAK activation and PP2A-mediated dephosphorylation of DLC1, which increases DLC1 GAP activity toward Rho GTPases, thereby suppressing cell spreading. In parallel, Rac1 activity in epidermal stem cells is modulated to balance migration and stasis during wound healing. DNA methylation patterns in CD4+ T-cells have been associated with psoriasis, suggesting epigenetic regulation of immune pathways that may influence epidermal wound healing.
negative regulation of wound healing, spreading of epidermal cells and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| RAC1 | Impaired wound healing, chronic wounds | Conditional knockout in epidermal stem cells; Rac1 inhibitor treatment |
| DLC1 | Cancer, metastasis; negative regulation of cell spreading | DLC1 knockout and overexpression in keratinocytes; EGF stimulation |
| CD4 | Psoriasis, psoriatic arthritis | DNA methylation profiling in CD4+ T-cells from patients |
| FAK (PTK2) | Wound healing, cancer | FAK knockout or point mutation in epidermal cells |
| PPP2CA | Wound healing, cancer | PP2A catalytic subunit knockout or knockdown |
Psoriasis and chronic inflammatory skin diseases
Psoriasis is a chronic inflammatory skin disease characterized by hyperproliferation and abnormal differentiation of epidermal cells. DNA methylation patterns in CD4+ T-cells can separate psoriasis patients from healthy controls and distinguish skin psoriasis from psoriatic arthritis, indicating that epigenetic and immune mechanisms contribute to disease pathogenesis. Dysregulation of negative regulatory pathways that normally restrain epidermal cell spreading may contribute to the epidermal hyperplasia seen in psoriasis.
Chronic wounds and impaired re-epithelialization
Chronic wounds fail to heal in a timely manner, often due to impaired epidermal cell migration and spreading. Rac1 activity in epidermal stem cells is required for efficient migration during wound healing, and its modulation can affect healing outcomes. Excessive negative regulation of epidermal spreading could contribute to delayed wound closure, making this pathway a potential therapeutic target.
Cancer and metastasis
The molecular machinery that controls epidermal cell spreading, including Rac1 and DLC1, is also implicated in cancer cell migration and metastasis. DLC1 is a known tumor suppressor whose GAP activity toward Rho GTPases can inhibit cell migration and invasion. Understanding how EGF/FAK/PP2A signaling activates DLC1 may provide insights into cancer progression and metastasis.
From negative regulation of wound healing, spreading of epidermal cells-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Is Rac1 required for epidermal stem cell migration during wound healing? | Conditional Rac1 knockout in mouse epidermis or human keratinocytes |
| Does DLC1 negatively regulate epidermal cell spreading? | DLC1 knockout and overexpression in keratinocytes, with EGF stimulation |
| How does EGF activate DLC1 through FAK and PP2A? | Point mutations in FAK or PP2A phosphorylation sites; knock-in of phospho-deficient DLC1 |
| What is the role of CD4+ T-cell methylation in psoriasis? | DNA methylation profiling and CRISPR editing of methylation sites in CD4+ T-cells |
| Can overexpression of DLC1 suppress epidermal hyperplasia? | Inducible DLC1 overexpression in mouse epidermis or 3D skin equivalents |
| What genes are essential for negative regulation of epidermal spreading? | Genome-wide CRISPR knockout library screening in keratinocytes |
How to Study the negative regulation of wound healing, spreading of epidermal cells Process
| Method | What It Measures | Typical Application |
|---|---|---|
| CRISPR knockout screen | Gene essentiality for cell spreading | Identify negative regulators in keratinocytes |
| Live-cell imaging | Dynamics of cell spreading and migration | Quantify Rac1 and DLC1 effects |
| Scratch wound assay | Rate of epidermal sheet closure | Assess wound healing in vitro |
| Phosphoproteomics | Phosphorylation changes downstream of EGF/FAK/PP2A | Map DLC1 regulatory sites |
| DNA methylation profiling | Epigenetic marks in CD4+ T-cells | Psoriasis biomarker discovery |
| Immunofluorescence | Localization of focal adhesion proteins | Assess cytoskeletal remodeling |
| Western blot | Protein expression and phosphorylation | Validate signaling pathways |
| qRT-PCR | mRNA expression levels | Measure gene expression changes |
CRISPR knockout screening
Genome-wide CRISPR knockout screens in epidermal keratinocytes can identify genes whose loss enhances or suppresses cell spreading. This approach is powerful for discovering novel negative regulators within GO:1903690.
Live-cell imaging and migration assays
Time-lapse microscopy of epidermal cells expressing fluorescently tagged cytoskeletal or focal adhesion markers allows quantification of spreading dynamics. Scratch wound assays and transwell migration assays measure the rate of epidermal sheet expansion.
Phosphoproteomics and signaling analysis
Mass spectrometry-based phosphoproteomics can map signaling events downstream of EGF, FAK, and PP2A, revealing how DLC1 phosphorylation regulates its GAP activity.
Epigenetic profiling
DNA methylation arrays or bisulfite sequencing of CD4+ T-cells can identify epigenetic changes associated with psoriasis, providing insights into immune-mediated regulation of epidermal wound healing.
How CRISPR Can Be Used to Study GO:1903690 negative regulation of wound healing, spreading of epidermal cells
Knockout
CRISPR knockout of candidate genes such as RAC1 or DLC1 in epidermal cells can test their requirement for negative regulation of wound healing. For example, Rac1 knockout in epidermal stem cells impairs migration, while DLC1 knockout may enhance spreading.
Point Mutation
CRISPR point mutation can introduce phospho-deficient or phospho-mimetic mutations in DLC1 or FAK to dissect signaling events downstream of EGF. This allows precise testing of phosphorylation sites identified by phosphoproteomics.
Knock-in
Knock-in of fluorescent tags or epitope tags into endogenous loci (e.g., DLC1-GFP) enables real-time imaging of protein localization and dynamics during epidermal cell spreading.
Overexpression
CRISPR activation (CRISPRa) or lentiviral overexpression of DLC1 or other negative regulators can test whether increased expression suppresses epidermal cell spreading and wound healing in vitro and in vivo.
How EDITGENE Supports negative regulation of wound healing, spreading of epidermal cells Research
Researchers studying negative regulation of wound healing, spreading of epidermal cells-related genes often need to determine whether a candidate gene is causally involved in restraining epidermal migration or whether it is merely correlated with the process. EDITGENE provides end-to-end CRISPR services to enable such causal experiments.
Contact EDITGENE today to design your custom CRISPR model for negative regulation of wound healing, spreading of epidermal cells research.
Frequently Asked Questions About negative regulation of wound healing, spreading of epidermal cells
What is GO:1903690?
GO:1903690 is a Gene Ontology biological process term defined as any process that stops, prevents, or reduces the frequency, rate, or extent of wound healing, spreading of epidermal cells.
What genes are involved in negative regulation of wound healing, spreading of epidermal cells?
Key genes include RAC1, DLC1, FAK (PTK2), PPP2CA, and EGF, which form a signaling axis controlling epidermal cell migration and spreading.
How does Rac1 regulate epidermal stem cell migration during wound healing?
Rac1 is a Rho-family GTPase that drives actin cytoskeletal reorganization and migration in epidermal stem cells; its modulation affects the rate of wound healing.
What is the role of DLC1 in epidermal cell spreading?
DLC1 is a Rho GTPase-activating protein that is activated by EGF via FAK and PP2A, leading to suppression of cell spreading.
How is EGF signaling linked to negative regulation of epidermal cell spreading?
EGF activates DLC1 through focal adhesion kinase (FAK) and protein phosphatase 2A (PP2A), which increases DLC1 GAP activity and inhibits cell spreading.
What diseases are associated with dysregulation of epidermal wound healing?
Psoriasis and chronic wounds are associated with dysregulation of epidermal wound healing; DNA methylation patterns in CD4+ T-cells distinguish psoriasis patients from healthy controls.
How can CRISPR be used to study GO:1903690?
CRISPR knockout, point mutation, knock-in, and overexpression can be used to test the causal role of candidate genes in negative regulation of epidermal cell spreading.
What methods are used to measure epidermal cell spreading?
Live-cell imaging, scratch wound assays, and transwell migration assays are commonly used to quantify epidermal cell spreading and migration.
What is the connection between psoriasis and epidermal wound healing?
Psoriasis involves abnormal epidermal proliferation and immune dysregulation; DNA methylation changes in CD4+ T-cells are associated with the disease, suggesting epigenetic links to epidermal wound healing pathways.
How can EDITGENE help my research on GO:1903690?
EDITGENE provides CRISPR knockout, point mutation, knock-in, overexpression, library screening, and bioinformatics services to study genes involved in negative regulation of wound healing, spreading of epidermal cells.
Conclusion
GO:1903690, negative regulation of wound healing, spreading of epidermal cells, represents a critical braking mechanism that ensures epidermal migration is properly controlled during wound repair. The Rac1 GTPase and the DLC1 Rho GTPase-activating protein, regulated by EGF/FAK/PP2A signaling, are central players in this process. Dysregulation of these pathways is linked to chronic wounds and inflammatory skin diseases such as psoriasis. CRISPR-based functional genomics, combined with imaging and proteomics, offers powerful tools to dissect the molecular mechanisms and identify new therapeutic targets within this pathway.
References
- 1. Chai LL et al.. 2011. [Modulatory effect of Rac1 protein on epidermal stem cells migration during wound healing].. Zhonghua Shao Shang Za Zhi 27(3):205-9 PMID: 21781463
- 2. Natoli V et al.. 2023. DNA methylation patterns in CD4(+) T-cells separate psoriasis patients from healthy controls, and skin psoriasis from psoriatic arthritis.. Front Immunol 14:1245876 PMID: 37662940
- 3. Ravi A et al.. 2015. Epidermal growth factor activates the Rho GTPase-activating protein (GAP) Deleted in Liver Cancer 1 via focal adhesion kinase and protein phosphatase 2A.. J Biol Chem 290(7):4149-62 PMID: 25525271
