• INHBE (Inhibin Subunit Beta E) is a member of the transforming growth factor-beta (TGF-β) superfamily and functions as a liver-enriched hepatokine involved in systemic metabolic regulation. INHBE expression is associated with lipid metabolism, insulin resistance, adipose tissue remodeling, and obesity-related metabolic disorders.
• Human genetic and translational studies have reported that elevated INHBE expression positively correlates with body mass index (BMI), insulin resistance, and hepatic metabolic dysfunction. Functional studies further indicate that inhibition of INHBE signaling can reduce body weight gain and adiposity, supporting its therapeutic potential in obesity and metabolic disease.
• INHBE humanized Mice were generated by replacing mouse Inhbe coding exons with human INHBE counterparts while retaining the endogenous mouse promoter and regulatory elements. This strategy enables physiologically relevant expression of human INHBE in vivo while disrupting endogenous mouse Inhbe expression.
• This model supports translational evaluation of INHBE-targeted RNAi therapeutics, antisense oligonucleotides (ASO), obesity drug discovery, metabolic disease studies, hepatokine biology research, and pharmacodynamic assessment of human-specific therapeutics.

Key Advantages

• Physiologically relevant human INHBE expression driven by endogenous mouse promoter
• Suitable for RNAi and antisense therapeutic evaluation
• Supports obesity and metabolic disease research
• Enables translational hepatokine biology studies
• Validated high-fat diet-induced obesity model
• Applicable for pharmacodynamic and efficacy studies of human-specific therapeutics

Validation

• Molecular Validation: Mouse Inhbe mRNA was detectable in wild-type C57BL/6 mice, whereas human INHBE mRNA was detectable in homozygous INHBE humanized Mice by RT-PCR and RT-qPCR.
• Protein Validation: Humanized INHBE protein expression was confirmed by western blot analysis in serum, liver, and kidney tissues of INHBE Humanized Mice.
• Metabolic Regulation Validation: Human INHBE expression was significantly increased under fasting conditions, consistent with the biological role of INHBE in lipid metabolism and suppression of lipolysis.
• Functional Validation: INHBE-targeting nucleic acid therapeutics significantly reduced human INHBE mRNA expression in vivo, supporting RNAi therapeutic evaluation of INHBE humanized Mice.
• Disease Model Validation: High-fat diet (HFD)-induced obesity studies demonstrated that INHBE-targeted siRNA reduced body weight gain, adipose tissue accumulation, hepatic lipid levels, and metabolic dysfunction.

Application

• RNAi and ASO therapeutic evaluation
• Obesity and metabolic disease research
• Hepatokine biology studies
• High-fat diet-induced obesity models
• Pharmacodynamic assessment of INHBE-targeted therapeutics
• Lipid metabolism and insulin resistance studies

INHBE

• The exons 1-2 of mouse Inhbe gene that encode the whole molecule (ATG to STOP codon), including 3’UTR were replaced by human counterparts in INHBE humanized mice. The promoter and 5’UTR region of the mouse gene are retained. The human INHBE expression is driven by endogenous mouse Inhbe promoter, while mouse Inhbe gene transcription and translation will be disrupted.

Strain-specific analysis of INHBE mRNA expression in wild-type C57BL/6JNifdc mice and INHBE humanized mice by RT-PCR. Liver RNA was isolated from wild-type C57BL/6JNifdc mice (+/+) and homozygous INHBE humanized mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human INHBE primers. Mouse Inhbe mRNA was detectable only in wild-type C57BL/6JNifdc mice. Human INHBE mRNA was detectable only in homozygous INHBE humanized mice but not in wild-type mice.

Western blot analysis of INHBE protein expression in homozygous INHBE humanized mice. Serum was collected from wild-type C57BL/6JNifdc mice (+/+) and homozygous INHBE humanized mice (H/H), and then analyzed by western blot with anti-INHBE antibody (Anti-Inhibin beta E chain antibody, ab103167). 20 μg (for INHBE) or 8 μg (for Transferrin) total proteins were loaded for western blot analysis. INHBE was detected in the serum of wild-type mice and homozygous INHBE humanized mice, as the antibody cross-recognizes both human and mouse INHBE.

Biochemical test of INHBE humanized mice. Blood biochemical parameters were evaluated in 7-week-old INHBE humanized mice. Values are expressed as mean ± SD.

The inhibitory efficiency of the nucleic acid drugs against human INHBE in INHBE humanized mice. INHBE humanized mice were randomly divided into two groups (male, 9-10 weeks old). The human INHBE targeted nucleic acid drugs (provided by a client), and PBS were administered to the mice individually. The nucleic acid drugs was administered in the form of PBS aqueous solution. The mice were sacrificed on day 5, and the liver tissue was collected to detect the expression level of human INHBE mRNA by qPCR. (A) The schematic diagram of experimental processing. (B) The expression of human INHBE mRNA in the liver. The human INHBE in the CPD-1 group was significantly reduced compared to the control group. Significance was determined by t-test, *P<0.05. Values are expressed as mean ± SEM.

The inhibitory efficiency of the nucleic acid drugs against human INHBE in INHBE humanized mice. INHBE humanized mice. were randomly divided into two groups (Male, 6-7 weeks old). The human INHBE targeted nucleic acid drugs (synthesized according to patents), and PBS were administered to the mice individually. The mice were sacrificed on day 14, and the liver tissue was collected to detect the expression level of human INHBE mRNA by qPCR. (A) The schematic diagram of experimental processing. (B) The expression of human INHBE mRNA in the liver. Significance was determined by t-test, *P<0.05. Values are expressed as mean ± SEM.

The inhibitory efficiency of the nucleic acid drugs against human INHBE in INHBE humanized mice and INHBE humanized mice plus. INHBE humanized mice and INHBE humanized mice plus were randomly divided into two groups (Male and female n=4/group). The human INHBE targeted nucleic acid drugs (provided by a client), and PBS were administered to the mice individually. The serum and liver tissue was collected after 6 hours of fasting to detect the expression level of human Activin E and INHBE mRNA. (A) The schematic diagram of experimental processing. (B) The expression of human Activin E in the serum on days 0, 7, 14, 21, and 28. (C) The expression of human INHBE mRNA in the liver. The human Activin E expression in the siRNA group was significantly reduced compared to the control group. Values are expressed as mean ± SEM.

High-Fat Diet-Induced INHBE humanized mice. C57BL/6JNifdc mice and INHBE humanized mice (male, 7 weeks old) were fed with high-fat diet (60 kcal% Fat) for 12 weeks to induce obesity. The human INHBE targeted nucleic acid drugs and PBS were administered to the mice individually.

High-Fat Diet-Induced INHBE humanized mice. C57BL/6JNifdc mice and INHBE humanized mice (male, 7 weeks old) were fed with a high-fat diet (60 kcal% Fat) for 12 weeks to induce obesity. (A) Body weight after HFD induction. (B) After fasting for 6 hours, mice were intraperitoneally injected with 15% Glucose (1.5 g/kg) for Glucose Tolerance Tests after 9 weeks of induction. (C) After fasting for 4 hours, mice were intraperitoneally injected with  0.5 U/kg insulin for the Insulin Tolerance Test after 10 weeks of induction.

Efficacy study of INHBE-siRNA AD-1708473.1 analog in HFD-induced INHBE humanized mice. (A) The expression of human INHBE mRNA in the liver at the end of the treatment. (B) Body weight changes after treatment. (C) Weight of white adipose tissue at the end of the treatment. (D-E) Hepatic TC and TG at the end of the treatment. Analyzed by one way-ANOVA, *P<0.05, **P<0.01, ***P<0.001. Values are expressed as mean ± SEM.

Efficacy study of INHBE-siRNA AD-1708473.1 analog in HFD-induced INHBE humanized mice. The 3D reconstruction analysis of lean tissue and adipose tissue by micro-CT after 6 times of treatment. (A-E) The weight of lean tissue, lean tissue which excludes internal organs, total adipose tissue, visceral adipose tissue, and subcutaneous adipose tissue. (F-J) The percentages of lean tissue, lean tissue which excludes internal organs, total adipose tissue, visceral adipose tissue, and subcutaneous adipose tissue with the mouse body weight.  Analyzed by one way-ANOVA, *P<0.05, **P<0.01, ***P<0.001.

High-Fat Diet-Induced INHBE humanized mice. C57BL/6JNifdc mice and INHBE humanized mice (male, 7 weeks old) were fed with high-fat diet (60 kcal% Fat) for 12 weeks to induce obesity. The human INHBE targeted nucleic acid drugs and PBS were administered to the mice individually.

Efficacy study of INHBE-siRNA in HFD-induced INHBE humanized mice. (A) The expression of human INHBE mRNA in the liver at the end of the treatment. (B) Body weight changes after treatment. (C) Weight of white adipose tissue at the end of the treatment. (D-E) Hepatic TC and TG at the end of the treatment. Analyzed by one way-ANOVA, *P<0.05, **P<0.01, ***P<0.001.

Efficacy study of INHBE-siRNA in HFD-induced INHBE humanized mice. The 3D reconstruction analysis of lean tissue and adipose tissue by micro-CT after 13 times of treatment. (A-E) The weight of lean tissue, lean tissue which excludes internal organs, total adipose tissue, visceral adipose tissue, and subcutaneous adipose tissue. (F-J) The percentages of lean tissue, lean tissue which excludes internal organs, total adipose tissue, visceral adipose tissue, and subcutaneous adipose tissue with the mouse body weight.  Analyzed by one way-ANOVA, *P<0.05, **P<0.01, ***P<0.001.

Efficacy study of INHBE-siRNA in HFD-induced INHBE humanized mice. The 3D reconstruction analysis of lean tissue and adipose tissue was performed by micro-CT after 13 treatments. (A-C) Representative images of the coronal plane by micro-CT. (D-F)  Representative images of 3D reconstruction by micro-CT.

Q1：What are INHBE humanized mice?

INHBE Humanized Mice are genetically engineered mice expressing human INHBE under endogenous regulatory control, enabling translational metabolic disease and RNAi therapeutic studies.

Q2: Why is INHBE an important therapeutic target?

INHBE is a hepatokine associated with obesity, insulin resistance, lipid metabolism, and adipose tissue regulation, making it a promising target for metabolic disease therapies.

Q3: Can these mice be used for siRNA evaluation?

Yes. The mRNA expression study demonstrated efficient in vivo knockdown of human INHBE using RNAi therapeutics.

Q4: Are these mice suitable for obesity studies?

Yes. INHBE humanized mice have been validated in high-fat diet-induced obesity models with metabolic and adipose tissue phenotyping.

Q5: What therapeutic modalities can be evaluated using this model?

This model supports evaluation of antisense oligonucleotides (ASO), RNAi therapeutics, and other human-specific INHBE-targeted interventions.