CTSG Antibody

What is Cathepsin G (CTSG) and why is it significant in immunological research?

Cathepsin G is an enzymatic protein belonging to the peptidase S1 protein family, encoded by the CTSG gene mapped to chromosome 14q12 in humans. This serine protease is primarily found in azurophil granules of neutrophilic polymorphonuclear leukocytes and exhibits specificity similar to chymotrypsin C. Its significance in immunological research stems from its dual functions: pathogen killing through proteolytic activity and participation in complex immunoregulatory processes .

CTSG contributes to pathogen elimination and tissue remodeling at inflammation sites, but also plays sophisticated roles in immune regulation. The protease participates in critical immune mechanisms including autoantigen processing, lymphocyte activation, and complement pathway modulation . This multifunctionality makes CTSG antibodies essential tools for investigating neutrophil function, inflammatory cascades, and immune dysregulation in various pathological states.

How do researchers distinguish between CTSG and other neutrophil serine proteases when designing experiments?

When designing experiments to specifically target CTSG while excluding other neutrophil serine proteases (NSPs) such as neutrophil elastase and proteinase 3, researchers should implement a multi-level experimental approach:

• Antibody selection strategy: Choose antibodies raised against unique epitopes of CTSG. For example, antibodies targeting positions R25-E252 of human CTSG provide good specificity as demonstrated in validated antibody products .

• Validation methodology: Always confirm antibody specificity through multiple techniques:

  • Western blot analysis showing the expected 29 kDa band for CTSG

  • Competitive binding assays with purified CTSG protein

  • Comparison with knockout/knockdown controls

• Functional discrimination: Incorporate activity-based assays that exploit CTSG's unique substrate preferences compared to other NSPs.

• Cross-reactivity assessment: Test potential cross-reactivity with other NSPs using purified proteins in parallel experiments to establish detection specificity .

The molecular weight verification is particularly important - CTSG appears at approximately 29 kDa on Western blots, distinguishing it from other NSPs .

What are the optimal protocols for using CTSG antibodies in Western blot applications?

For optimal Western blot detection of CTSG, researchers should follow this methodologically rigorous protocol based on validated research approaches:

Sample Preparation:

• Prepare neutrophil-rich samples or cell lines known to express CTSG (e.g., THP-1, MCF-7)

• Use appropriate lysis buffer containing protease inhibitors excluding serine protease inhibitors when studying active CTSG

• Load 20-30 μg of protein per lane for optimal detection

Electrophoresis Conditions:

• Employ 5-20% gradient SDS-PAGE gel

• Run at 70V (stacking gel)/90V (resolving gel) for 2-3 hours

• Include molecular weight markers capable of identifying the expected 29 kDa band

Transfer and Blocking:

• Transfer to nitrocellulose membrane at 150 mA for 50-90 minutes

• Block with 5% non-fat milk in TBS for 1.5 hours at room temperature

Antibody Incubation:

• Primary antibody: Dilute anti-CTSG antibody to 0.1-0.5 μg/ml in blocking buffer

• Incubate overnight at 4°C

• Wash with TBS-0.1% Tween 3 times, 5 minutes each

• Secondary antibody: Anti-rabbit IgG-HRP at 1:5000 dilution for 1.5 hours at room temperature

Detection:

• Develop using enhanced chemiluminescence (ECL) system

• Expected band for CTSG appears at approximately 29 kDa

This protocol has been validated with cell lines including human THP-1 and MCF-7, providing consistent detection of the target protein .

How should researchers optimize immunohistochemistry procedures when using CTSG antibodies?

For optimal immunohistochemical detection of CTSG in tissue sections, researchers should implement the following methodological approach:

Tissue Preparation:

• Fix tissues in 10% neutral buffered formalin

• Embed in paraffin and section at 4-6 μm thickness

• Mount on positively charged slides

Antigen Retrieval (Critical Step):

• Perform heat-mediated antigen retrieval using EDTA buffer (pH 8.0)

• Heat to 95-100°C for 15-20 minutes followed by cooling to room temperature

Staining Protocol:

• Block endogenous peroxidase with 3% H₂O₂ for 10 minutes

• Block non-specific binding with 10% goat serum for 30-60 minutes

• Incubate with anti-CTSG antibody at 2-5 μg/ml concentration overnight at 4°C

• Apply peroxidase-conjugated secondary antibody (goat anti-rabbit IgG) for 30 minutes at 37°C

• Develop with DAB chromogen and counterstain with hematoxylin

Validation Controls:

• Positive control: Human tonsil tissue (demonstrates robust CTSG expression)

• Negative controls: Primary antibody omission and isotype controls

Evaluation Parameters:

• Assess cellular location of staining (should be cytoplasmic granular pattern in neutrophils)

• Quantify staining intensity and distribution (0-3+ scoring system)

• Document neutrophil infiltration patterns around areas of interest

This protocol has been validated on human tonsil tissue, showing specific labeling of neutrophil populations containing CTSG .

What considerations are important when analyzing CTSG expression in inflammatory and autoimmune disease models?

When analyzing CTSG expression in inflammatory and autoimmune disease models, researchers must account for several critical methodological factors:

Sample Timing and Disease Phase:

• Collect samples at multiple time points to capture dynamic expression changes

• Differentiate between acute and chronic phases of inflammation

• Consider diurnal variations in neutrophil activity and CTSG release

Comprehensive Expression Analysis:

• Measure both mRNA and protein levels (may not correlate due to post-transcriptional regulation)

• Assess CTSG activity using specific enzymatic assays

• Correlate with clinical biomarkers (e.g., creatine kinase and lactate dehydrogenase in dermatomyositis)

Cellular Context Evaluation:

• Distinguish between intracellular and extracellular CTSG

• Assess neutrophil extracellular trap (NET) formation and CTSG association

• Evaluate CTSG binding to cell surfaces vs. soluble forms

Disease-Specific Considerations:

• For dermatomyositis: Correlate CTSG activity with Jo-1 auto-antibody status

• For multiple sclerosis: Assess CTSG's effect on myelin basic protein degradation

• For type 1 diabetes: Measure CTSG-mediated proinsulin processing and T cell activation

Analytical Framework:

• Use cell-specific markers to identify CTSG-expressing populations

• Implement multiplexed assays to assess CTSG alongside related inflammatory mediators

• Consider spatial distribution using imaging techniques

Research shows that CTSG mRNA and activity levels are elevated in autoimmune conditions like dermatomyositis, multiple sclerosis, and type 1 diabetes mellitus, making these considerations essential for accurate interpretation of disease mechanisms .

How can researchers effectively investigate CTSG's role in antigen processing and T cell activation?

To effectively investigate CTSG's role in antigen processing and T cell activation, researchers should implement a comprehensive experimental framework:

Antigen Processing Analysis:

• In vitro degradation assays:

  • Incubate purified CTSG with candidate autoantigens (e.g., myelin basic protein, proinsulin)

  • Use mass spectrometry to identify specific cleavage sites and resulting peptide fragments

  • Compare digestion patterns with and without specific CTSG inhibitors

• Epitope mapping:

  • Determine if CTSG-generated fragments contain known T cell epitopes

  • Assess epitope binding to MHC class II molecules using binding assays

  • Example: CTSG degrades the immunodominant MBP epitope (MBP85-99) and generates another T cell epitope (MBP115-123)

T Cell Activation Studies:

• Functional T cell assays:

  • Use CTSG-processed antigens to stimulate antigen-specific T cells

  • Measure proliferation, cytokine production, and activation markers

  • Compare responses to unprocessed antigens and antigens processed by other proteases

• Mechanistic investigations:

  • Assess if CTSG activates T cells through direct binding to lymphocyte receptors

  • Evaluate CTSG's effect on both Th1 and Th2 pathways

  • Research shows CTSG augments antigen-specific antibody production via T cell activation

Inhibition Studies:

• Use selective CTSG inhibitors to confirm specificity of observed effects

• Implement CTSG knockdown/knockout approaches in relevant cell types

• Assess downstream consequences on antigen presentation and T cell responses

Translational Approach:

• For type 1 diabetes: Study how CTSG processes proinsulin into intermediates that polarize T cell activation

• For multiple sclerosis: Investigate how CTSG-mediated degradation of myelin basic protein impacts T cell responses

• Monitor changes in CD4+ T cell activation following CTSG inhibition

Research data demonstrates that CTSG plays crucial roles in processing key autoantigens such as proinsulin and myelin basic protein, making it a central player in autoimmune disease pathogenesis .

What methodological approaches can determine CTSG's contribution to vascular permeability in inflammation?

To determine CTSG's contribution to vascular permeability in inflammation, researchers should implement these methodological approaches:

In Vitro Endothelial Barrier Function Assays:

• Transendothelial electrical resistance (TEER) measurements:

  • Culture human dermal microvascular endothelial cells on semipermeable membranes

  • Apply purified CTSG at physiologically relevant concentrations

  • Monitor real-time changes in electrical resistance as indicator of barrier integrity

  • Include specific CTSG inhibitors as controls

• Endothelial permeability assays:

  • Measure passage of labeled dextrans or albumin across endothelial monolayers

  • Quantify permeability coefficients with and without CTSG treatment

  • Assess dose-response relationships and kinetics of barrier disruption

Molecular Mechanism Studies:

• Receptor-mediated effects:

  • Evaluate CTSG-induced expression of protease-activated receptor 2 (PAR2) on endothelial cells

  • Use PAR2 antagonists to determine receptor dependency

  • Assess downstream signaling pathways (Ca²⁺ flux, MAPK activation)

• Cytoskeletal alterations:

  • Visualize F-actin reorganization using fluorescence microscopy

  • Quantify focal adhesion density and distribution

  • Monitor VE-cadherin and tight junction protein localization

In Vivo Vascular Leakage Models:

• Intravital microscopy:

  • Administer fluorescent tracers and observe extravasation in real-time

  • Apply CTSG locally and monitor vascular leakage

  • Use CTSG knockout models or inhibitors for validation

• Modified Miles assay:

  • Inject Evans blue dye systemically

  • Apply CTSG to skin or other tissues

  • Quantify dye extravasation spectrophotometrically

Clinical Correlation Studies:

• Measure circulating CTSG levels in patients with inflammatory conditions

• Correlate with markers of endothelial dysfunction and clinical parameters

• Assess vascular leakage using imaging techniques

Research data shows that CTSG can induce expression of protease-activated receptor 2 and alter the cytoskeleton of human dermal microvascular endothelial cells, thereby contributing to increased lymphocyte infiltration in inflammatory conditions like dermatomyositis .

How can researchers explore CTSG as a potential biomarker in cancer progression?

To explore CTSG as a potential biomarker in cancer progression, researchers should employ a systematic investigative approach:

Expression Analysis in Cancer Tissues:

• Multi-omics profiling:

  • Analyze CTSG mRNA expression across cancer types using RNA-seq data

  • Perform proteomic analysis to quantify CTSG protein levels

  • Integrate with genomic data to identify correlations with mutations or copy number variations

  • Example: Bioinformatics analyses from TCGA data identified CTSG as a potential biomarker in oral squamous cell carcinoma

• Tissue microarray analysis:

  • Create tissue microarrays from cancer and matched normal tissues

  • Perform immunohistochemistry using validated CTSG antibodies

  • Quantify expression using digital pathology approaches

  • Correlate with clinicopathological parameters

Functional Validation Studies:

• Cell line models:

  • Manipulate CTSG expression in cancer cell lines using overexpression/knockdown

  • Assess effects on proliferation, migration, invasion, and colony formation

  • Evaluate changes in immune cell recruitment and tumor microenvironment

• Patient-derived xenograft models:

  • Establish PDX models from tumors with varying CTSG expression

  • Monitor tumor growth rates and metastatic potential

  • Test CTSG inhibitors for therapeutic potential

Biomarker Validation Framework:

• Discovery phase:

  • Use bioinformatics to identify CTSG from immune-related gene datasets

  • Create co-expression networks with transcription factors

  • Perform survival analysis to correlate expression with outcomes

• Validation phase:

  • Confirm findings in independent patient cohorts

  • Develop standardized assays for clinical application

  • Assess CTSG in liquid biopsies (serum, plasma)

Clinical Implementation Research:

• Determine optimal cutoff values for prognostic stratification

• Integrate with existing biomarker panels

• Evaluate predictive value for treatment response

• Conduct longitudinal studies to assess dynamic changes during treatment

This methodological framework is supported by research indicating that CTSG has potential as a prognostic biomarker derived from immune-related genes in oral squamous cell carcinoma, as identified through bioinformatics analyses of RNA-seq data from The Cancer Genome Atlas .

What are the most common technical challenges when working with CTSG antibodies and how can they be overcome?

Researchers frequently encounter several technical challenges when working with CTSG antibodies. Here are the most common issues and methodological solutions:

Low Signal Intensity:

• Problem: Insufficient detection of CTSG despite known expression

  • Solution: Optimize protein extraction using specialized buffers for granular proteins

  • Methodology: Include mild detergents (0.5-1% NP-40) in lysis buffer and avoid freeze-thaw cycles

  • Validation: Compare fresh vs. stored samples to establish optimal processing protocols

• Problem: Ineffective antigen retrieval in fixed tissues

  • Solution: Test multiple antigen retrieval methods sequentially

  • Methodology: Compare citrate buffer (pH 6.0) vs. EDTA buffer (pH 8.0) and adjust heating time

  • Evidence-based approach: Research shows EDTA buffer (pH 8.0) provides optimal retrieval for CTSG in paraffin-embedded sections

Non-specific Binding:

• Problem: High background or multiple bands in Western blot

  • Solution: Implement stringent blocking and washing protocols

  • Methodology: Block with 5% non-fat milk/TBS for 1.5 hours, then use 0.1% Tween in TBS for washing

  • Optimization parameter: Adjust antibody concentration to 0.1-0.5 μg/ml for optimal signal-to-noise ratio

• Problem: Non-specific staining in IHC

  • Solution: Include additional blocking steps and controls

  • Methodology: Block with 10% goat serum, add avidin/biotin blocking for biotin-based detection systems

  • Validation controls: Include isotype controls and tissue known to be negative for CTSG

Sample Degradation:

• Problem: CTSG activity loss during sample processing

  • Solution: Implement specialized sample handling protocols

  • Methodology: Process samples immediately or stabilize with appropriate inhibitors

  • Technical approach: Store in buffer containing 4 mg Trehalose, 0.9 mg NaCl, and 0.2 mg Na₂HPO₄

Antibody Storage and Stability:

• Problem: Loss of antibody activity during storage

  • Solution: Follow optimized storage conditions

  • Methodology: Store lyophilized antibody at -20°C; after reconstitution, store at 4°C for one month or aliquot and freeze at -20°C for six months

  • Technical advice: Avoid repeated freeze-thaw cycles to maintain antibody performance

These solutions have been validated across multiple experimental systems, ensuring reliable detection of CTSG in various research applications.

How can researchers distinguish between active and inactive forms of CTSG in experimental settings?

Distinguishing between active and inactive forms of CTSG requires specialized techniques that detect conformational and functional differences. Here's a comprehensive methodological approach:

Activity-Based Protein Profiling:

• Mechanism: Uses activity-based probes that covalently bind only to catalytically active CTSG

  • Methodology: Incubate samples with fluorescent/biotinylated serine protease activity probes

  • Analysis: Detect labeled active CTSG via gel electrophoresis or fluorescence imaging

  • Validation: Include known inhibitors of CTSG to confirm specificity

Enzymatic Activity Assays:

• Chromogenic/fluorogenic substrate assays:

  • Methodology: Use CTSG-specific substrates (e.g., Suc-Ala-Ala-Pro-Phe-pNA)

  • Quantification: Measure absorbance/fluorescence changes over time

  • Controls: Compare to standard curves using purified active CTSG

• Zymography:

  • Methodology: Incorporate CTSG substrates into non-reducing SDS-PAGE gels

  • Analysis: Identify active CTSG as cleared bands after Coomassie staining

  • Technical enhancement: Combine with Western blotting for dual detection of total and active CTSG

Immunological Approaches:

• Conformation-specific antibodies:

  • Methodology: Use antibodies that selectively recognize active site conformations

  • Application: Apply in immunoblotting, ELISA, or imaging techniques

  • Validation: Confirm with recombinant active vs. inactive CTSG controls

• Immunoprecipitation-activity assays:

  • Methodology: Immunoprecipitate CTSG with non-inhibitory antibodies, then measure activity

  • Analysis: Calculate ratio of activity to total protein as indication of activation state

Cellular Localization Studies:

• Subcellular fractionation:

  • Methodology: Separate granular, cytosolic, and membrane fractions

  • Analysis: Active CTSG relocates from granules to membranes or extracellular space upon activation

  • Technical approach: Compare distribution across fractions using both activity assays and immunoblotting

In Situ Approaches:

• FRET-based reporters:

  • Methodology: Design FRET sensors containing CTSG-specific cleavage sequences

  • Analysis: Measure FRET signal changes in live cells or tissues

  • Application: Enables real-time spatial monitoring of CTSG activation

These approaches provide complementary information about CTSG's activation state, critical for understanding its roles in physiological and pathological processes such as immune cell activation and autoimmune disease progression .

What emerging technologies might enhance CTSG research in autoimmune and inflammatory diseases?

Several cutting-edge technologies are poised to revolutionize CTSG research in autoimmune and inflammatory diseases over the next decade:

Single-Cell Multi-omics Approaches:

• Single-cell proteogenomics:

  • Simultaneously profile CTSG gene expression and protein levels at single-cell resolution

  • Identify previously uncharacterized CTSG-expressing cell populations in disease tissues

  • Map heterogeneity in CTSG activation states across immune cell subpopulations

• Spatial transcriptomics and proteomics:

  • Visualize CTSG expression patterns within the tissue microenvironment

  • Correlate spatial distribution of CTSG with immune cell infiltration and tissue damage

  • Create 3D maps of CTSG activity gradients in inflammatory lesions

Advanced Imaging Technologies:

• Intravital multiphoton microscopy with CTSG activity reporters:

  • Monitor CTSG activity in real-time within living tissues

  • Track neutrophil behavior and CTSG release during inflammation

  • Assess immediate effects of CTSG inhibitors on immune cell dynamics

• Super-resolution microscopy:

  • Visualize subcellular localization of CTSG at nanometer resolution

  • Study CTSG trafficking between cellular compartments

  • Examine CTSG incorporation into neutrophil extracellular traps (NETs)

CRISPR-Based Functional Genomics:

• Cell-type specific CTSG knockouts:

  • Generate conditional CTSG knockouts in specific neutrophil populations

  • Study tissue-specific roles of CTSG in autoimmune models

  • Create humanized mouse models expressing human CTSG variants

• High-throughput CRISPR screens:

  • Identify genetic modifiers of CTSG expression and activity

  • Discover novel regulators in the CTSG pathway

  • Map synthetic lethal interactions for therapeutic targeting

AI and Machine Learning Applications:

• Predictive modeling of CTSG involvement in disease progression:

  • Integrate multi-omics data to predict CTSG-dependent disease outcomes

  • Develop algorithms to identify patients likely to benefit from CTSG-targeted therapies

  • Create decision support tools for precision medicine approaches

Therapeutic Development Platforms:

• Selective CTSG inhibitors with improved pharmacokinetics:

  • Design tissue-specific drug delivery systems for CTSG inhibitors

  • Develop dual-action inhibitors targeting CTSG and complementary pathways

  • Creating monitoring systems for tissue-specific CTSG inhibition

Research in type 1 diabetes has already demonstrated that CTSG inhibitors can reduce blood glucose levels, improve islet beta cell function, and reduce CD4+ T cell activation, suggesting significant therapeutic potential for these emerging approaches .

How might computational approaches enhance understanding of CTSG's role in disease networks?

Computational approaches offer powerful methods to enhance understanding of CTSG's role in complex disease networks. Here are key methodological frameworks researchers can implement:

Network Biology and Systems Medicine:

• Protein-protein interaction (PPI) network analysis:

  • Construct CTSG-centered interactomes using experimental and predicted interactions

  • Identify hub proteins and network motifs connecting CTSG to disease pathways

  • Apply graph theory to quantify network perturbations in disease states

• Multi-scale modeling of inflammatory cascades:

  • Develop mathematical models integrating CTSG enzymatic kinetics with cellular responses

  • Simulate temporal dynamics of CTSG-mediated inflammation

  • Predict system-level consequences of CTSG inhibition

Advanced Bioinformatics Approaches:

• Gene regulatory network inference:

  • Identify transcription factors regulating CTSG expression

  • Map enhancers and silencers controlling tissue-specific CTSG levels

  • Create co-expression networks associating CTSG with disease-relevant gene modules

  • Example: Co-expression network analysis identified CTSG as a potential biomarker in oral squamous cell carcinoma

• Pathway enrichment and functional annotation:

  • Apply ontology-based enrichment to identify biological processes linked to CTSG

  • Perform cross-disease analysis to find common CTSG-dependent mechanisms

  • Integrate proteomics data to map post-translational modification landscapes

Structure-Based Computational Methods:

• Molecular dynamics simulations:

  • Model CTSG conformational dynamics in different microenvironments

  • Simulate interactions with substrates and inhibitors

  • Predict effects of disease-associated mutations on CTSG function

• Virtual screening and drug repurposing:

  • Screen compound libraries for novel CTSG inhibitors

  • Identify FDA-approved drugs with CTSG-modulating potential

  • Design peptide-based inhibitors targeting specific CTSG functions

Machine Learning Applications:

• Predictive biomarker discovery:

  • Develop algorithms to identify patients with CTSG-driven disease subtypes

  • Create classifiers predicting response to CTSG-targeted therapies

  • Build integrated risk scores incorporating CTSG activity with clinical parameters

• Natural language processing of biomedical literature:

  • Extract CTSG-related knowledge from published research

  • Identify understudied connections between CTSG and disease mechanisms

  • Generate testable hypotheses for experimental validation

Translational Bioinformatics:

• Electronic health record mining:

  • Correlate CTSG biomarker levels with clinical outcomes

  • Identify phenotypic patterns associated with altered CTSG function

  • Assess real-world effectiveness of interventions targeting CTSG pathways

These computational approaches can significantly accelerate research by identifying new connections between CTSG and disease mechanisms, prioritizing therapeutic targets, and enabling precision medicine strategies for conditions where CTSG plays a pivotal role.

What standardized protocols should researchers follow when validating and characterizing new CTSG antibodies?

Researchers should follow these standardized protocols when validating and characterizing new CTSG antibodies:

Table 1: Comprehensive CTSG Antibody Validation Protocol

Detailed Application-Specific Protocols:

• Western Blot Validation:

  • Run standard SDS-PAGE with 5-20% gradient gels

  • Transfer to nitrocellulose membrane at 150 mA for 50-90 minutes

  • Block with 5% non-fat milk/TBS for 1.5 hours

  • Incubate with antibody at 0.1-0.5 μg/ml overnight at 4°C

  • Use anti-rabbit IgG-HRP at 1:5000 and develop with ECL

• Immunohistochemistry Validation:

  • Perform heat-mediated antigen retrieval in EDTA buffer (pH 8.0)

  • Block with 10% goat serum

  • Incubate with antibody at 2-5 μg/ml overnight at 4°C

  • Apply peroxidase-conjugated secondary antibody for 30 minutes

  • Develop with DAB and counterstain with hematoxylin

• Functional Inhibition Testing:

  • Pre-incubate CTSG with antibody at various ratios

  • Measure residual enzymatic activity using specific substrates

  • Calculate IC50 values to quantify inhibitory potency

• Epitope Mapping:

  • Generate peptide arrays covering CTSG sequence

  • Probe with antibody to identify binding regions

  • Confirm accessibility of epitope in native protein structure

The validation should include positive controls such as human THP-1 and MCF-7 whole cell lysates for Western blotting and human tonsil tissue for immunohistochemistry, as these have been established as reliable sources of CTSG expression .

What are the key experimental parameters when studying CTSG in different disease models?

When studying CTSG in different disease models, researchers should consider these key experimental parameters:

Table 2: Critical Parameters for CTSG Analysis in Disease Models

Disease-Specific Methodological Considerations:

• Dermatomyositis:

  • Measure CTSG mRNA in peripheral blood mononuclear cells and muscle tissue

  • Correlate serum CTSG activity with creatine kinase and lactate dehydrogenase

  • Stratify analysis based on Jo-1 auto-antibody status

  • Assess CTSG-induced protease activated receptor 2 expression on endothelial cells

• Multiple Sclerosis:

  • Analyze CTSG-mediated degradation of myelin basic protein

  • Focus on destruction of immunodominant epitopes (e.g., MBP85-99)

  • Evaluate generation of new T cell epitopes (e.g., MBP115-123)

  • Test binding of processed peptides to MHC class II molecules

• Type 1 Diabetes:

  • Assess CTSG-mediated proinsulin degradation and processing

  • Measure activation of proinsulin-reactive T cells

  • Monitor blood glucose levels, islet β-cell function, and CD4+ T cell activation

  • Test CTSG inhibitors' effects on disease progression

• Oral Squamous Cell Carcinoma:

  • Analyze RNA-seq data for CTSG expression patterns

  • Create co-expression networks with immune-related genes and transcription factors

  • Perform survival analysis based on CTSG expression levels

  • Integrate with clinical parameters for prognostic modeling

Methodological Framework for Cross-Disease Comparisons:

• Standardize sample collection and processing protocols

• Employ consistent CTSG detection and activity measurement methods

• Use matched controls appropriate for each disease context

• Apply uniform statistical analyses to enable cross-disease comparisons

• Consider tissue-specific and species-specific variations in CTSG expression and function

This structured approach enhances reproducibility and facilitates meaningful comparisons of CTSG's roles across different disease contexts, providing a comprehensive understanding of its pathophysiological significance .