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Anti-HER2 CAR-T Preclinical in vivo Assay

Target Background

Human epidermal growth factor receptor 2 (HER2) belongs to transmembrane epidermal growth factor receptor family and encoded by gene ERBB2. There is no ligand reported to directly activate HER2 and HER2 can heterodimerize with other family member receptors such as HER1, HER3 and HER4. Homo- or heterodimerization activation of HER2 lead to phosphorylation of intracellular tyrosine residue and stimulate various signaling pathways including MAPK, PI3K and PKC, which results in cell proliferation, differentiation, angiogenesis and migration. HER2 has been identified as a valuable tumor-associated target for cancer therapy. Overexpression of HER2 is reported in breast cancer patient, as well as other forms of cancer including stomach, ovary, uterine and brain cancers.

Anti-HER2 CAR-T Preclinical in vivo Assay

Engineering CAR-T cells: Design concepts.
Trends in Immunology,2015. 36(8), 494-502.

Anti-HER2 CAR-T Cell Therapy

Several Phase I clinical trials are being conducted to evaluate the safe dose and adverse effects of anti-HER2 CAR-T cells for patients with breast cancer, sarcoma and glioblastoma (NCT02442297, NCT00902044 and NCT02547961). Completed phase I/II studies indicate that HER2-specific CAR-T cells can persist for 6 weeks in patients of osteosarcomas without dose-limiting toxicity.

Animal Models for in vivo Study of anti-HER2 CAR-T Cell Therapy

Creative Biolabs provides various xenograft tumor models to support anti-HER2 CAR-T therapy research. Our technical team has extensive knowledge and experience for in vivo model development. All animal experiments are monitored under an approved Institutional Animal Care and Use Committee (IACUC) protocol.
Xenograft model of human breast cancer
Immunodeficient mice received subcutaneously injection of human breast cancer cells SKBR3 on right or left flank. When tumor expansion meet criterion, anti-HER2 CAR-T cells are injected intravenously into mice. Tumor size are measured by calipers and volume are calculated by using reported formula.
Xenograft model of human osteosarcoma
Loco-regional model is developed by intraperitoneal injection of LM7 cell and followed by HER2-specific CAR-T cells treatment. As for systemic model, recipient mice are anesthetized with inhalation isoflurane and followed by tail-vein injection of LM7 cells or luciferase expressing LM7 cells, and effector cells.
Xenograft model of brain tumor
For human medulloblastoma and glioblastoma model, NOD-SCID mice are bred and prepared for tumor implantation by anesthetizing with isofluorane (or intraperitoneally injection of Avertin solution). Head of mice are shaved and fixed in adaptor. Medulloblastoma cells Daoy or GBM cells U373 are transplanted through 1 mm burr-hole into skull. CAR-T cells are treated in the same coordinates, which locates in the center of right caudate nucleus. Incision is closed after effector cells injection by sutures. Pain control like buprenorphine can be used if necessary.

In vivo Assay Parameters and Techniques

Creative Biolabs offers comprehensive services to test anti-HER2 CAR-T therapy in preclinical trials. All tests are conduct by well-trained technicians and strike to milestone of timeline.
Efficacy Tests
Tumor remission monitored by tumor volume recording or bioluminescence imaging and survival curve tracking.
Viability and Bio-distribution Studies
Durability and bio-distribution are evaluated by bioluminescence imaging, immunochemistry staining and real-time PCR
Toxicity Evaluation
Pilot tolerated evaluation: route of administration, dosage, MTD
Clinical observation: body weight, food consumption, behavior and pathological signs
Cytokine storm surveillance (fever, hypertension, prolonged cytopenia)
Postmortem analysis
Tumorigenicity study
GLP-Compliant Preclinical Test
All our experiments are performed by well-trained and experienced technicians in a GLP-compliant and IACUC-regulated facility.

Creative Biolabs fully understands researches of adoptive T cell therapy targeting HER2 are time-consuming and costly. With our highly experienced technical groups and state-of-the-art facilities, Creative Biolabs would like to provide well-proved in vivo tumor models to accelerate the process of CAR-T cell therapy evaluation.


  1. Ahmed, et al. "HER2-specific T cells target primary glioblastoma stem cells and induce regression of autologous experimental tumors." Clinical Cancer Research 16.2 (2010): 474-485.
  2. Ahmed, et al. "Immunotherapy for osteosarcoma: genetic modification of T cells overcomes low levels of tumor antigen expression." Molecular Therapy 17.10 (2009): 1779-1787.
  3. Iqbal, et al. "Human epidermal growth factor receptor 2 (HER2) in cancers: overexpression and therapeutic implications." Molecular biology international 2014 (2014).

All services and products are only for lab research use, not for any clinical diagnosis or treatment.

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