CCDC23 Human

What is CCDC23 and what are its primary cellular functions? (Basic)

CCDC23, also known as Small vasohibin-binding protein (SVBP), is a human protein consisting of 66 amino acids that plays several critical roles in cellular function. Its primary functions include:

• Enhancement of the tyrosine carboxypeptidase activity of VASH1 and VASH2, promoting the removal of C-terminal tyrosine residues from alpha-tubulin

• Regulation of microtubule dynamics critical for spindle function and chromosome segregation during mitosis

• Enhancement of the solubility and secretion of VASH1 and VASH2 proteins

• Participation in axon and excitatory synapse formation in neuronal cells

The protein's sequence (MGSSHHHHHHSSGLVPRGSHM GSEFMDPPARKEKTKVKESVS RVEKAKQKSAQQELKQRQRAEI YALNRVMTELEQQQFDEFCKQM QPPGE) contains structural elements that facilitate its interaction with binding partners, particularly vasohibin proteins .

How is CCDC23 expression altered during aging? (Basic)

Research on gene expression profiles in blood cells has identified CCDC23 as one of only four genes that are dysregulated more than 2-fold with aging. Specifically, CCDC23 was found to be significantly up-regulated in elderly individuals compared to adults. This finding emerged from a study comparing healthy adults (average age 46±3 years) with elderly individuals (average age 68±4 years) .

The upregulation of CCDC23 with age suggests it may play a role in age-related cellular changes, potentially affecting microtubule dynamics and neuronal function in aging populations. Though all parameters in the study subjects remained within normal physiological ranges, the consistent upregulation indicates CCDC23 may be a biological marker of aging processes .

What methodological approaches are used to study CCDC23 protein interactions? (Basic)

Several complementary methodological approaches are employed to investigate CCDC23 protein interactions, particularly with its known binding partners VASH1 and VASH2:

• Recombinant protein production: Expression of CCDC23 in Escherichia coli systems with appropriate purification tags (such as the His-tag system described in search result ) to obtain >90% purity for downstream applications

• Co-immunoprecipitation assays: To confirm physical interactions between CCDC23 and target proteins in cellular contexts

• SDS-PAGE and Western blotting: To analyze protein expression and interaction patterns

• Mass spectrometry (MS): For detailed characterization of protein complexes and post-translational modifications

• Functional enzymatic assays: To measure how CCDC23 enhances the tyrosine carboxypeptidase activity of vasohibin proteins

These methodologies provide complementary data on both the physical interactions and functional consequences of CCDC23 binding to partner proteins .

How should experiments be designed to investigate CCDC23's role in microtubule dynamics? (Advanced)

When designing experiments to study CCDC23's role in microtubule dynamics, researchers should implement a comprehensive approach that addresses both molecular mechanisms and cellular consequences:

In vitro experimental approaches:

• Purified protein assays using recombinant CCDC23 (>90% purity) with VASH1/2 and tubulin substrates

• Quantitative assessment of tubulin detyrosination activity with and without CCDC23

• Microtubule polymerization/depolymerization kinetics measurements

• Structural studies of CCDC23-VASH complexes bound to tubulin substrates

Cellular experimental approaches:

• CRISPR-mediated CCDC23 knockout or siRNA knockdown in relevant cell types

• Live-cell imaging of fluorescently-tagged tubulin to track dynamic instability parameters

• Immunofluorescence analysis of post-translationally modified tubulin populations

• Mitotic spindle analysis in cells with altered CCDC23 expression

Control considerations:

• Comparison of VASH1/2 activity with and without CCDC23 co-expression

• Use of catalytically inactive VASH mutants as negative controls

• Rescue experiments reintroducing wild-type or mutant CCDC23

• Synchronized cell populations to control for cell cycle variables

Proper experimental design requires rigorous controls, appropriate randomization, and sufficient statistical power as outlined in standard experimental design guidelines .

What considerations are important when interpreting contradictory findings in CCDC23 studies? (Advanced)

When faced with contradictory findings in CCDC23 functional studies, researchers should employ a structured approach to interpretation:

Methodological reconciliation:

• Compare experimental systems used (cell lines, tissues, in vitro assays)

• Evaluate differences in protein expression levels and purification methods

• Assess sensitivity and specificity of detection methods

• Consider whether the same functional domains or protein isoforms were studied

Statistical considerations:

• Evaluate sample sizes and statistical power across studies

• Assess whether appropriate statistical tests were applied

• Consider potential sources of variability in the experimental system

• Determine if multiple testing corrections were properly applied when appropriate

Experimental validation approaches:

• Design experiments that directly test competing hypotheses

• Utilize multiple complementary methods to address the same question

• Perform independent validation in different model systems

• Consider context-dependent functions based on cell type or physiological state

This systematic approach helps distinguish technical artifacts from genuine biological complexity while advancing understanding of CCDC23 function following principles of good experimental design practice .

How can researchers ensure ethical compliance when designing human subject research involving CCDC23? (Basic)

Research involving human subjects that investigates CCDC23 must adhere to established ethical standards and regulatory requirements:

Regulatory compliance:

• All research involving human subjects conducted or supported by institutions must comply with established protection policies (such as HHS Policy for Protection of Human Research Subjects, 45 CFR Part 46)

• Research must be approved by an institutional review board (IRB) prior to initiation

• Investigators must maintain competency in research ethics and human research regulations through appropriate training

Experimental design considerations:

• Clear definition of variables related to CCDC23 expression or function that will be measured

• Specific, testable hypotheses about CCDC23's role in the research question

• Proper assignment of subjects to treatment groups with appropriate controls

• Adequate study size to ensure statistical power while minimizing unnecessary subject recruitment

Historical context awareness:

• Familiarity with foundational ethical guidelines including the Belmont Report, Declaration of Helsinki, and Nuremberg Code

• Application of established principles for protection of human subjects

• Consideration of international ethical guidelines when conducting research across different regions

Researchers should maintain communication with their institutional IRB throughout the research process and contact appropriate authorities (such as huma@cdc.gov) for additional guidance on human subjects protection .

What experimental models are most appropriate for studying CCDC23 function in neurodevelopment? (Advanced)

Given CCDC23's role in axon and excitatory synapse formation, selecting appropriate experimental models for neurodevelopmental studies is critical:

Cellular models:

• Primary neuronal cultures from rodent embryonic cortex or hippocampus

• Human induced pluripotent stem cell (iPSC)-derived neurons

• 3D cerebral organoids that recapitulate aspects of human brain development

• Neuroblastoma cell lines for initial mechanism studies

In vivo models:

• Conditional CCDC23 knockout mice with neuron-specific Cre drivers

• In utero electroporation for spatiotemporal control of CCDC23 expression

• Zebrafish models for high-throughput developmental studies

• Drosophila models for genetic interaction studies

Analytical approaches:

• Time-lapse imaging of neurite outgrowth and branching

• Quantitative assessment of synapse density and morphology

• Electrophysiological recordings to correlate structure with function

• Molecular analysis of tubulin post-translational modifications

When designing these experiments, researchers must consider developmental timing, regional specificity, and cell-type specific effects while maintaining appropriate controls to isolate CCDC23-specific phenotypes from potential compensatory mechanisms .

What statistical approaches are most appropriate for analyzing CCDC23 expression changes in aging studies? (Advanced)

Based on findings that CCDC23 is significantly upregulated during aging, appropriate statistical frameworks for analyzing such expression changes include:

Experimental design considerations:

• Age as continuous vs. categorical variable

• Longitudinal vs. cross-sectional study design

• Sample size determination through power analysis

• Controlling for potential confounding variables (sex, health status, medication use)

Statistical analysis framework:

Validation approaches:

• qPCR validation of expression changes with careful reference gene selection

• Protein-level confirmation through Western blotting or proteomics

• Cellular phenotype correlation with expression levels

• Meta-analysis across multiple aging studies

These approaches were successfully applied in the referenced aging study that identified CCDC23 as one of four genes dysregulated more than 2-fold with aging .

What are the current challenges and solutions in producing functional recombinant CCDC23 for structural studies? (Advanced)

Production of functional recombinant CCDC23 for structural and interaction studies presents several challenges that can be addressed through optimized protocols:

Expression system selection:

• E. coli expression: Successfully used to produce full-length human CCDC23 (aa 1-66) with >90% purity as demonstrated in published protocols

• Fusion tags: His-tag systems (MGSSHHHHHHSSGLVPRGSHM) have been effectively employed for affinity purification

• Expression conditions: Optimization of induction parameters, temperature, and media composition to maximize soluble protein yield

Purification challenges and solutions:

• Solubility: Co-expression with binding partners (VASH1/VASH2) may enhance solubility

• Stability: Buffer optimization to maintain protein integrity during purification

• Purity assessment: SDS-PAGE and mass spectrometry verification of >90% purity for downstream applications

Functional validation:

• Activity assays to confirm that recombinant CCDC23 enhances vasohibin enzymatic function

• Binding assays to verify interaction with known partners

• Circular dichroism to assess proper protein folding

The published protocol achieving >90% purity of recombinant human CCDC23 expressed in E. coli provides a validated starting point for researchers seeking to conduct structural and functional studies .

How does CCDC23 dysregulation contribute to age-related cellular phenotypes? (Advanced)

The significant upregulation of CCDC23 in elderly individuals suggests several potential mechanisms by which it may contribute to age-related cellular phenotypes:

Microtubule dynamics alterations:

• Increased CCDC23 levels may enhance vasohibin-mediated tubulin detyrosination

• Altered post-translational modification patterns could affect microtubule stability

• Changes in microtubule dynamics may influence cellular processes including division, transport, and migration

• Potential impacts on mitotic fidelity and chromosomal stability in aging cells

Neuronal implications:

• Given CCDC23's role in axon and excitatory synapse formation, upregulation may alter neuronal connectivity

• Changes in synaptic maintenance may contribute to age-related cognitive changes

• Altered microtubule dynamics could affect neuronal transport and signaling

Cellular stress response:

• CCDC23 upregulation might represent a compensatory mechanism to counteract other age-related cellular stresses

• Integration with other age-related gene expression changes suggests potential involvement in broader aging networks

The aging study identified CCDC23 as one of only four genes dysregulated more than 2-fold with aging, highlighting its potential significance in age-related biological processes .

What methodological approaches can distinguish between correlation and causation in studies linking CCDC23 to disease? (Advanced)

Establishing causal relationships between CCDC23 and disease processes requires rigorous methodological approaches:

Experimental strategies:

• Genetic manipulation:

  • CRISPR/Cas9-mediated knockout or knockdown in disease models

  • Overexpression studies to mimic disease-associated upregulation

  • Knock-in of disease-associated variants

  • Rescue experiments with wild-type CCDC23

• Mechanistic studies:

  • Identification of complete molecular pathways linking CCDC23 to disease phenotypes

  • Intervention at multiple pathway points to test necessity and sufficiency

  • Temporal manipulation of CCDC23 at different disease stages

Study design considerations:

• Human studies:

  • Prospective cohort studies tracking CCDC23 expression before disease onset

  • Case-control studies with appropriate matching and confounding control

  • Genetic association studies if CCDC23 variants are identified

• Experimental design principles:

  • Clear definition of variables (CCDC23 as independent variable)

  • Appropriate control groups

  • Minimization of confounding variables

  • Adequate sample size for statistical power

These approaches must adhere to ethical standards for human subjects research, including IRB approval and proper informed consent procedures .

How can emerging technologies advance CCDC23 functional characterization? (Basic)

Several cutting-edge technologies offer promising approaches to deepen our understanding of CCDC23 function:

Advanced imaging technologies:

• Super-resolution microscopy to visualize CCDC23 localization at nanometer resolution

• Live-cell imaging combined with optogenetic tools for temporal control of CCDC23 function

• Correlative light and electron microscopy to connect CCDC23 function with ultrastructural features

Genetic and genomic technologies:

• CRISPR screening approaches to identify genetic interactors

• Single-cell transcriptomics to identify cell populations with unique CCDC23 expression patterns

• Spatial transcriptomics to map CCDC23 expression in complex tissues

Protein analysis technologies:

• Hydrogen-deuterium exchange mass spectrometry to study CCDC23 dynamics

• Cryo-electron microscopy for high-resolution structures of CCDC23-vasohibin complexes

• Proximity labeling approaches to map the CCDC23 interactome in living cells

These technologies will enable researchers to address fundamental questions about CCDC23 function, regulation, and its contributions to both normal physiology and disease states.

What are the most promising therapeutic approaches targeting the CCDC23 pathway? (Advanced)

While direct therapeutic targeting of CCDC23 remains in early research stages, several approaches show potential based on its molecular function:

Direct targeting strategies:

• Small molecule modulators of CCDC23-vasohibin interactions

• Peptide-based inhibitors that disrupt specific protein-protein interfaces

• RNA-based therapeutics (siRNA, antisense oligonucleotides) for temporary modulation

Indirect targeting approaches:

• Modulation of enzymes affected by CCDC23 activity

• Targeting downstream effectors in the microtubule modification pathway

• Cell type-specific delivery systems for CNS applications given CCDC23's role in neurons

Therapeutic contexts:

• Age-related disorders given CCDC23's upregulation in aging

• Neurodevelopmental conditions based on its role in axon and synapse formation

• Cell division disorders linked to its function in mitotic spindle regulation

Development considerations:

• Target validation through genetic models

• Biomarker development to identify responsive populations

• Combination approaches with existing therapeutics

• Ethical considerations regarding human subjects protection in clinical trials

Any therapeutic development program would need to adhere to established protocols for human subjects protection, including appropriate IRB oversight and informed consent procedures .