• CD3 is composed of four protein chains, including CD3E, CD3D, CD3G, and CD3Z, which serve as critical components of the TCR/CD3 complex on T cells. The CD3 complex plays essential roles in T cell antigen recognition, signal transduction, immune activation, and T cell development.
• Cadherin 17 (CDH17) is a member of the 7D-cadherin superfamily characterized by seven cadherin repeats and a short cytoplasmic domain. CDH17 is primarily expressed in the gastrointestinal tract, especially in the small intestine and colon, where it contributes to epithelial integrity, intercellular adhesion, peptide transport, and calcium-dependent water absorption.
• In CD3EDG/CDH17 Humanized Mice, chimeric human CD3EDG genes were engineered while endogenous mouse Cd3edg genes are disrupted. In addition, the extracellular domain of mouse Cdh17 was replaced with the corresponding human CDH17 extracellular region, enabling evaluation of human-specific therapeutics targeting CDH17.
• Human CD3E was detected on T cells of homozygous CD3EDG/CDH17 Humanized Mice, while human CDH17 expression was confirmed in colon and small intestine tissues.
• This model is suitable for preclinical pharmacodynamic and toxicity studies of bispecific antibody-drug conjugates (BsADCs), T cell engagers, and other immunotherapies targeting CDH17-positive solid tumors, including colorectal cancer and gastrointestinal cancers.

Key Advantages

• Dual humanized CD3EDG and CDH17 platform
• Suitable for T cell engager and BsADC evaluation
• Supports gastrointestinal cancer immunotherapy studies
• Humanized CD3 signaling for translational immune activation studies
• Humanized CDH17 expression in gastrointestinal tissues
• Applicable for efficacy and toxicity assessment of human-specific therapeutics

Validation

• Molecular Validation: Mouse Cd3e, Cd3d, Cd3g, and Cdh17 mRNA were detectable only in wild-type C57BL/6JNifdc mice, whereas human CD3E, CD3D, CD3G, and CDH17 mRNA were detectable in homozygous CD3EDG/CDH17 Humanized Mice by RT-PCR.
• Protein Validation: Human CD3E protein was detectable on splenic and peripheral blood T cells of homozygous CD3EDG/CDH17 Humanized Mice by flow cytometry.
• Tissue Validation: Humanized CDH17 protein expression was confirmed in colon and small intestine tissues by western blot analysis.
• Histopathological Validation: IHC analysis demonstrated CDH17 expression patterns in gastrointestinal tissues of CD3EDG/CDH17 Humanized Mice.

Application

• Bispecific antibody-drug conjugate evaluation
• T cell engager efficacy studies
• Colorectal cancer research
• Gastrointestinal cancer immunotherapy
• CDH17-targeted therapeutic development
• Preclinical toxicity and pharmacodynamic studies

Strain-specific analysis of human CD3EDG/CDH17 mRNA expression was performed in wild-type C57BL/6JNifdc mice and CD3EDG/CDH17 Humanized Mice by RT-PCR. Colon RNA was isolated from wild-type C57BL/6JNifdc mice (+/+) and homozygous CD3EDG/CDH17 Humanized Mice (H/H;H/H). cDNA libraries were synthesized by reverse transcription followed by PCR using mouse- or human-specific CD3E, CD3D, CD3G, and CDH17 primers. Mouse Cd3e, Cd3d, Cd3g, and Cdh17 mRNA were detectable only in wild-type C57BL/6JNifdc mice. Human CD3E, CD3D, CD3G, and CDH17 mRNA were detectable in homozygous CD3EDG/CDH17 Humanized Mice but not in wild-type mice.

Strain-specific human CD3E expression analysis was performed in homozygous CD3EDG/CDH17 Humanized Mice by flow cytometry. Splenocytes (A) and blood cells (B) were collected from wild-type C57BL/6JNifdc mice (+/+) and homozygous CD3EDG/CDH17 Humanized Mice (H/H;H/H). Protein expression was analyzed using species-specific anti-mouse CD3E antibody (BioLegend, 100312) and anti-human CD3E antibody (BD Horizon™, 562426). Mouse CD3E was detectable in wild-type C57BL/6JNifdc mice. Human CD3E was detectable on T cells of homozygous CD3EDG/CDH17 Humanized Mice but not in wild-type mice.

Western blot analysis of human CDH17 protein expression was performed in homozygous CD3EDG/CDH17 Humanized Mice. Colon and small intestine tissue lysates were collected from wild-type C57BL/6JNifdc mice (+/+) and homozygous CD3EDG/CDH17 Humanized Mice (H/H;H/H) and analyzed by western blot using species-specific anti-human CDH17 antibody (Abcam, ab109190) and anti-mouse CDH17 antibody (R&D Systems, AF8524). A total of 40 μg protein was loaded for western blot analysis. CDH17 protein was detected in colon and small intestine tissues.

Immunohistochemical (IHC) analysis of human CDH17 expression was performed in wild-type C57BL/6JNifdc mice and homozygous CD3EDG/CDH17 Humanized Mice. Various tissues were collected and analyzed by IHC using anti-CDH17 antibody (Abcam, ab109190).
CDH17 was detectable in both wild-type mice and homozygous CD3EDG/CDH17 Humanized Mice because of antibody cross-reactivity between mouse and human CDH17.

Q1：What are CD3EDG/CDH17 Humanized Mice?

CD3EDG/CDH17 Humanized Mice are genetically engineered mice expressing humanized CD3EDG and CDH17 targets for translational immunotherapy and cancer research.

Q2: Why is CDH17 an important therapeutic target?

CDH17 is highly expressed in gastrointestinal cancers and plays important roles in epithelial integrity and tumor biology, making it a promising target for cancer therapeutics.

Q3: Can these mice be used for bispecific antibody evaluation?

Yes. The model is specifically designed for evaluation of BsADCs, T cell engagers, and other human-specific CDH17-targeted therapeutics.

Q4: What tissues express humanized CDH17 in this model?

Humanized CDH17 expression was confirmed in colon and small intestine tissues.

Q5: Are these mice suitable for toxicity studies?

Yes. The model supports both efficacy and toxicity assessment of CDH17-targeted immunotherapies in vivo.