Endometrial Cancer

• Ishikawa: one of the most classic models in low-grade, differentiated, and estrogen-dependent endometrial cancer research, suitable for hormonal therapy, progesterone research, ER/PR pathway, PI3K/AKT/mTOR pathway, metabolism-related drugs, and long-term maintenance therapy research.
• Ishikawa-luc: suitable for orthotopic uterine models, long-term hormonal intervention, withdrawal rechallenge, and local relapse dynamic monitoring.
• Ishikawa models require a clear hormone background; if the program involves estrogen, progesterone, or withdrawal rechallenge, hormone supplementation, withdrawal window, and long-term follow-up should be set in the protocol design.

• HEC-1-A: suitable for invasive, non-hormone-dependent, chemotherapy combination, local infiltration, and partial MMR abnormality background research.
• HEC-1-B: suitable for invasion enhancement, dMMR/pMMR stratification exploration, peritoneal dissemination, and high-invasion program early validation; HEC-1-B-luc can be used for peritoneal dissemination and orthotopic model dynamic monitoring.
• HEC-1-A and HEC-1-B can be used for basic efficacy and PK/PD research under stratified background, but conventional immunodeficient mice cannot fully evaluate immunotherapy efficacy.

• KLE: commonly used for high-invasion, p53 abnormality, chemotherapy resistance, serous-like tendency, and relapsed/refractory direction research.
• AN3 CA: suitable for local invasion, orthotopic expansion, peritoneal dissemination, and advanced relapse research; AN3 CA-luc can be used for uterine orthotopic, peritoneal dissemination, and relapse model dynamic tracking.
• KLE and AN3 CA are more suitable for refractory and relapse scenarios, but should not represent low-grade, hormone-dependent populations.

• HER2 Targeted Programs: HER2 is enriched in serous-like, p53-abnormal, or high-risk subgroups. Confirm baseline target expression in candidate cell lines prior to model selection.
• ADC Programs: Select cell lines based on target antigen density, membrane localization, internalization kinetics, and in vivo retention. Prioritize HER2-positive, highly invasive cell lines for HER2-ADCs.
• Bispecifics, TCE & Cell Therapies: Verify target antigen expression in cell lines first, then advance to huPBMC or huHSC humanized platforms to validate human immune-mediated efficacy.

Subcutaneous CDX typically involves inoculating human endometrial cancer cells subcutaneously into immunodeficient mice, followed by randomization and dosing once tumors reach a predetermined volume. Commonly used hosts include BALB/c nude, NOD-SCID, and NSG mice.

• Differentiated and hormone-dependent subcutaneous CDX: Ishikawa, with estrogen or progesterone background as needed
• dMMR and invasive subcutaneous CDX: HEC-1-A, HEC-1-B
• High-invasion and serous-like subcutaneous CDX: KLE, AN3 CA

Model characteristics
Ishikawa is suitable for hormonal therapy, progesterone research, PI3K/AKT/mTOR inhibitors, metabolic agents, and long-term maintenance regimen early assessment. HEC-1-A or HEC-1-B are suitable for chemotherapy, combination targeted agents, dMMR/pMMR stratification background basic efficacy comparison, and local invasion direction validation. KLE or AN3 CA are suitable for high-invasion, advanced relapse, serous-like tendency, HER2, ADC, and chemotherapy resistance direction early screening. This model has high standardization and good reproducibility, suitable for early screening and PK/PD research; its limitations are the lack of uterine orthotopic microenvironment, myometrial invasion, peritoneal dissemination, and postoperative relapse processes.

Modeling and dosing timelines
Conventional subcutaneous inoculation typically reaches enrollment volume within 1–4 weeks, with randomization commonly performed at tumor volumes of approximately 80–150 mm³. The conventional treatment period is 2–4 weeks. For hormone resistance, withdrawal rechallenge, long-term maintenance, ADC interval dosing, or immune combination pre-treatment studies, this can be extended to 4–8 weeks. ER/PR, MMR, p53, HER2, and target antigen programs should re-verify key marker expression before modeling, at enrollment, and at the endpoint stage.

Detection endpoints

• Basic efficacy layer: TGI, RTV, terminal tumor weight, body weight, general condition, and progression-free time
• Molecular stratification layer: ER, PR, MMR, p53, HER2, PD-L1, Ki67, and target antigen expression
• Mechanism layer: Cleaved caspase-3, TUNEL, pAKT, pS6, CD31, apoptosis, cell cycle, ADC tissue distribution, and target pathway inhibition
• PK/PD layer: Plasma drug concentration, tumor tissue concentration, target occupancy, pathway inhibition, ADC distribution, ctDNA exploration, and exposure-efficacy relationship

Recommended positive controls
Carboplatin at 30–60 mg/kg IP or IV, once weekly or cyclically; paclitaxel at 10–20 mg/kg IP or IV, once weekly or cyclically. Medroxyprogesterone acetate, megestrol acetate, or other progestin agents can serve as hormone direction references. PI3K, AKT, or mTOR pathway programs can set pathway inhibitors as mechanism references. HER2-positive programs can set trastuzumab or trastuzumab deruxtecan as references.

Human orthotopic models typically use Ishikawa-luc, HEC-1-A-luc, AN3 CA-luc, or KLE-luc, inoculated or implanted into the uterine horn, uterine cavity-related sites, or local uterine tissue of NSG or NOD-SCID mice. Orthotopic burden can be tracked via BLI/IVIS, ultrasound, or terminal dissection.

Model characteristics
Ishikawa-luc is more suitable for hormone dependence, ER/PR, progesterone response, and long-term maintenance therapy research. HEC-1-A-luc is suitable for invasive and relapse directions; AN3 CA-luc and KLE-luc are more suitable for high-invasion, myometrial invasion, serous-like tendency, and peritoneal extension research. This model is closer to the real organ environment, suitable for local invasion, relapse, and long-term maintenance research; its limitations are complex surgery, lower throughput, and greater inter-batch variability.

Modeling and dosing timelines
Orthotopic inoculation typically forms monitorable burden within 1–4 weeks, and luc-labeled models can be stratified for enrollment via BLI. The conventional treatment period is 2–6 weeks. If studying hormone withdrawal, relapse, long-term maintenance, or local therapy, the observation period can be extended. Treatment can begin after orthotopic BLI signal confirmation, or after orthotopic burden reaches a predetermined standard, followed by withdrawal, induction therapy, or local treatment, then relapse observation.

Detection endpoints

• Orthotopic burden layer: BLI/IVIS, orthotopic tumor burden, uterine weight, uterine local pathology, survival, and body weight
• Local invasion layer: Myometrial invasion, uterine cavity lesions, periuterine tissue invasion, local relapse, and tissue pathology scoring
• Molecular mechanism layer: ER, PR, MMR, p53, HER2, Ki67, cleaved caspase-3, pAKT, pS6, and PD-L1
• Immune layer: In syngeneic or humanized models, CD8/Treg, MDSC, macrophages, cytokines, TCR repertoire, single-cell sequencing, and spatial immune analysis can be detected

Recommended positive controls
Ishikawa orthotopic models can set progestin agents as references. Carboplatin combined with paclitaxel can serve as an orthotopic model basic chemotherapy reference. dMMR/MSI-H or immune-related programs can reference pembrolizumab, dostarlimab, or durvalumab-related clinical pathways. HER2-positive high-invasion orthotopic models can set trastuzumab or trastuzumab deruxtecan references.

Peritoneal dissemination models typically use AN3 CA-luc, KLE-luc, or HEC-1-B-luc, established via intraperitoneal injection, or by spontaneous progression from high-invasion orthotopic models. NSG or NOD-SCID mice are commonly used hosts.

Model characteristics
Suitable for advanced dissemination, peritoneal implantation, post-relapse progression, high invasion, local peritoneal therapy, and anti-dissemination strategy research. Peritoneal dissemination models are suitable for high-invasion and advanced relapse scenarios, but have complex quantification and greater inter-batch variability, requiring unified BLI, nodule counting, and endpoint sampling standards.

Detection endpoints
Total peritoneal BLI signal, peritoneal nodule count, peritoneal burden score, body weight, survival, ascites exploration, and peritoneal lesion pathology.

Recommended positive controls
Carboplatin combined with paclitaxel as a basic reference; high-invasion or HER2 directions can set trastuzumab or trastuzumab deruxtecan based on target; immune-related programs should enter immune-compatible models.

Can use tail vein injection, left ventricle injection, intrasplenic injection, or portal vein injection to establish lung, liver, or other organ colonization models. Commonly used models include AN3 CA-luc, KLE-luc, HEC-1-B-luc, and high-invasion derivatives.

Model characteristics
Suitable for specific organ colonization, systemic dissemination, anti-metastasis agents, and advanced high-invasion mechanism research. Experimental distant metastasis models are typically not the main platform in endometrial cancer, and are more suitable for answering specific organ colonization questions. If the program focuses on local invasion and relapse, orthotopic models have higher priority.

Detection endpoints
Organ BLI, metastatic nodule count, organ weight, tissue pathology, survival, metastasis-related markers, and drug tissue exposure.

Recommended positive controls
Carboplatin combined with paclitaxel can serve as a systemic therapy reference. HER2-positive or target-positive programs should select corresponding target-positive models and verify metastatic lesion target expression. Immune-related programs should enter immune-compatible models.

Use AN3 CA-luc or KLE-luc to establish uterine orthotopic models, observing peritoneal dissemination or distant colonization after local progression. Induction therapy, withdrawal, or local treatment follow-up can be added as needed.

Model characteristics
Suitable for orthotopic invasion to dissemination full-process, relapse metastasis, long-term maintenance, and MRD clearance research. This model has stronger clinical relevance but longer cycles and greater variability, with higher requirements for orthotopic modeling stability and long-term tracking.

Detection endpoints
Orthotopic burden, peritoneal dissemination, distant colonization, BLI, survival, time to relapse, and pathology.

Recommended positive controls
Carboplatin combined with paclitaxel as a basic reference. HER2-positive or target-positive programs can set corresponding targeted agent references. Syngeneic orthotopic relapse models can set anti-PD-1, anti-PD-L1, or immune combination strategies.

Syngeneic subcutaneous models can use murine endometrial cancer syngeneic models or engineered syngeneic models in C57BL/6, BALB/c, or matched immunocompetent mice. Grouping occurs when tumors reach approximately 50–100 mm³.

Model characteristics
Suitable for immune checkpoint, STING, cytokines, radiotherapy combination, local therapy, and TME early screening.

Detection endpoints
Tumor volume, TGI, body weight, survival, flow cytometry immune profiling, CD8/Treg, myeloid cell profile, PD-L1, cytokines, and IHC/IF.

Recommended positive controls
Anti-PD-1 antibody, anti-PD-L1 antibody, anti-CTLA-4 antibody, STING agonists, or cytokine strategies can serve as immunotherapy references. Antibody doses can start at 100–200 μg/mouse IP, twice weekly or once every 3 days.

Syngeneic orthotopic uterine models can inoculate murine endometrial cancer cells or engineered cells into the uterine-related sites of matched strains. This model simultaneously retains the uterine local microenvironment and intact immune system.

Model characteristics
Suitable for dMMR immunotherapy, local therapy, radiotherapy combination, organ microenvironment, relapse, and immune microenvironment remodeling research. Syngeneic immunocompetent models can evaluate efficacy and mechanisms under an intact immune system, particularly suitable for immunotherapy and TME research. Their limitation is the difference between murine models and human endometrial cancer in molecular stratification and target expression.

Detection endpoints
Orthotopic burden, survival, local immune infiltration, CD8/Treg, MDSC, macrophages, PD-L1, cytokines, TCR repertoire, single-cell sequencing, and spatial immune analysis.

Recommended positive controls
Anti-PD-1 antibody, anti-PD-L1 antibody, anti-CTLA-4 antibody, STING agonists, or cytokine strategies can serve as immunotherapy references. Antibody doses can start at 100–200 μg/mouse IP, twice weekly or once every 3 days.

The huPBMC model establishes a short-term human immune effector environment by infusing human peripheral blood mononuclear cells into immunodeficient mice, combined with Ishikawa, HEC-1-A, HEC-1-B, KLE, or AN3 CA human endometrial cancer models.

Model characteristics
Suitable for bispecifics, TCE, CAR-T/NK, human cytokines, human immune checkpoints, and ADC or antibody programs highly dependent on human immune effects. This model has high cost, long cycles, significant donor variability, and carries xGvHD or cytokine release risk.

Detection endpoints
hCD45, hCD3, hCD4, hCD8, hNK, T cell activation, tumor infiltration, IFN-γ, TNF-α, IL-6, Granzyme B, tumor burden, body weight, CRS, and xGvHD risk markers.

Recommended positive controls
Anti-human PD-1 antibody, anti-human PD-L1 antibody, or anti-human CTLA-4 antibody can serve as mechanism references. TCE, CAR-T, or CAR-NK programs should include target-positive, low/negative, effector cell, and empty vector/irrelevant TCE controls. Cell therapy doses are often designed on the order of 10⁶–10⁷ cells/mouse.

The huHSC model reconstructs a longer-term human immune system via human hematopoietic stem cells, suitable for long-cycle human immune reconstitution, complex combination therapy, and immune resistance research.

Model characteristics
Suitable for sustained immune activation, bispecifics, TCE, CAR-T/NK, immune resistance, long-term maintenance therapy, and complex TME mechanism validation. This model has high cost, long cycles, significant donor variability, and carries xGvHD or cytokine release risk; it is not recommended as a first-round high-throughput screening platform.

Detection endpoints
Human immune reconstitution ratio, T/B/NK/myeloid cell spectrum, tumor infiltration, cytokines, human immune checkpoint expression, tumor burden, survival, and major organ pathology.

Recommended positive controls
Anti-human PD-1 antibody, anti-human PD-L1 antibody, or anti-human CTLA-4 antibody can serve as mechanism references. TCE, CAR-T, or CAR-NK programs should include target-positive, low/negative, effector cell, and empty vector/irrelevant TCE controls. Cell therapy doses are often designed on the order of 10⁶–10⁷ cells/mouse.

Relapse and MRD models are typically based on Ishikawa-luc, HEC-1-A-luc, AN3 CA-luc, or KLE-luc orthotopic uterine models. First, trackable orthotopic lesions are established, followed by induction therapy, hormone withdrawal, local treatment, or withdrawal observation. Progestins, carboplatin combined with paclitaxel, targeted agents, or immune combination strategies can be used to induce lesion burden reduction, then entering withdrawal or maintenance phase, observing BLI relapse, local relapse, peritoneal dissemination, and survival.

Model characteristics
Ishikawa is more suitable for hormone withdrawal and long-term maintenance; HEC-1-A is suitable for invasive relapse; AN3 CA and KLE are more suitable for high-invasion, serous-like, p53-abnormal, and peritoneal dissemination relapse. This model is suitable for maintenance therapy, relapse inhibition, MRD clearance, and long-term survival research, but is technically complex with longer cycles, requiring high demands for orthotopic tumor formation, local treatment timing, and long-term follow-up; it is not suitable for early large-scale screening.

Modeling and dosing timelines
First establish orthotopic uterine lesions and confirm burden by BLI/IVIS or imaging. Premature treatment may result in an insufficient relapse window, while delayed treatment may result in an excessively high dissemination baseline. Progestins, chemotherapy, targeted agents, or immune combination regimens are used to reduce or stabilize orthotopic burden, then entering maintenance/withdrawal phase. After withdrawal or de-escalation, observation continues for 4–10 weeks or longer, recording BLI relapse, local relapse, peritoneal dissemination, distant metastasis, and survival.

Detection endpoints

• MRD and relapse layer: BLI relapse, time to relapse, local relapse rate, peritoneal dissemination, distant colonization, survival, and ctDNA exploration
• Mechanism layer: ER, PR, MMR, p53, HER2, Ki67, cleaved caspase-3, pAKT, pS6, CD31, and target retention
• Immune layer: In syngeneic or humanized models, CD8/Treg, MDSC, macrophages, hCD45, cytokines, and TCR repertoire can be detected
• Safety layer: Body weight, uterine local pathology, liver and kidney function, complete blood count, major organ pathology, and dosing tolerance

Recommended positive controls
Ishikawa relapse models can set progestin maintenance therapy references. Carboplatin combined with paclitaxel can serve as a general chemotherapy relapse reference. HER2-positive or target-positive relapse models can set trastuzumab or trastuzumab deruxtecan references. Syngeneic orthotopic relapse models can set anti-PD-1, anti-PD-L1, or immune combination strategies.