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Heterogeneity Mapping of Protein Expression in Tumors using Quantitative Immunofluorescence
Published on: October 25, 2011
Intratumoral heterogeneity impacts the response to anti-neu antibody therapy
Hyunkeun Song, Tae Oh Kim, Sun Young Ma
1Departmentof Microbiology and Immunology, INJE University College of Medicine, 633-165 GaegumDong, Busanjin Gu, Busan 614-735, Republic of Korea. yxfu@bsd.uchicago.edu.
Background:
Along with de novo resistance, continued exposure to trastuzumab, an anti-human epidermal growth factor receptor 2 (HER2/neu) antibody, can lead to acquired resistance. In this study, we characterize a new anti-HER2/neu antibody resistant and metastatic mouse breast carcinoma cell line, TUBO-P2J. This cell line was developed during in vivo experiments using the antibody sensitive and non-metastatic tumor line TUBO. In addition, TUBO-P2J was used to establish an intratumoral HER2 heterogenous animal tumor model to evaluate the therapeutic effects of anti-HER2/neu antibody.
Methods:
After establishing the cell line, TUBO-P2J was characterized regarding its susceptibility to anti-neu antibody and chemotherapeutics, as well as its metastatic potential in vitro and in vivo. In addition, expression profiles of metastasis related genes were also evaluated. A clinically relevant intratumoral HER2 heterogenous tumor model was established by inoculating mice with tumor cells consisting of TUBO and TUBO-P2J at a ratio of 1,000:1 or 10,000:1. Tumor growth and mouse survival were used to evaluate the therapeutic effects of anti-neu antibody.
Results:
The TUBO-P2J cell line is a HER2/neu negative and highly metastatic variant of TUBO. This cell line was resistant to anti-neu antibody therapy, and when inoculated subcutaneously, metastasized to the lungs within 14 days. Compared to the parental TUBO cell line, TUBO-P2J displayed an epithelial-mesenchymal transition (EMT) related gene expression profile including: the loss of E-cadherin, and increased Vimentin, Snail, and Twist1 expression. In addition, TUBO-P2J exhibited increased invasion and migration activity, and was resistant to chemotherapy drugs. Finally, mixed tumor implantations experiments revealed that an increased percentage of TUBO-P2J rendered tumors less responsive to anti-neu antibody therapy.
Conclusion:
This study describes a novel model of intratumoral heterogenous metastatic breast cancer in immune competent mice that can be used to develop novel or combined immunotherapies to overcome antibody resistance.
Insights
A new metastatic breast cancer cell line, TUBO-P2J, developed resistance to anti-HER2/neu antibody therapy. This model helps study antibody resistance and develop new immunotherapies for HER2-positive breast cancer.
Area of Science:
- Oncology
- Immunotherapy
- Cancer Biology
Background:
- Trastuzumab, an anti-HER2/neu antibody, faces challenges with de novo and acquired resistance in breast cancer treatment.
- A new metastatic mouse breast carcinoma cell line, TUBO-P2J, was developed from the antibody-sensitive TUBO line to study resistance mechanisms.
Purpose of the Study:
- To characterize the TUBO-P2J cell line for its resistance to anti-HER2/neu antibodies and chemotherapeutics.
- To establish a clinically relevant intratumoral HER2 heterogeneous tumor model for evaluating therapeutic strategies.
- To investigate the metastatic potential and gene expression profiles associated with resistance.
Main Methods:
- TUBO-P2J cells were characterized for drug susceptibility, metastatic potential (in vitro and in vivo), and gene expression.
- An intratumoral HER2 heterogeneous model was created by co-inoculating TUBO and TUBO-P2J cells at varying ratios.
- Tumor growth and survival were monitored to assess the efficacy of anti-HER2/neu antibody therapy.
Main Results:
- TUBO-P2J is a HER2/neu negative, highly metastatic variant of TUBO, resistant to anti-HER2/neu antibody therapy.
- TUBO-P2J cells exhibited an epithelial-mesenchymal transition (EMT) gene expression profile, increased invasion, migration, and chemotherapy resistance.
- Tumors with a higher percentage of TUBO-P2J cells showed reduced responsiveness to anti-HER2/neu antibody treatment.
Conclusions:
- A novel model of intratumoral heterogeneous metastatic breast cancer in immunocompetent mice was established.
- This model can be utilized to develop and test novel or combined immunotherapies aimed at overcoming antibody resistance.
- The findings provide a platform for advancing therapeutic strategies in HER2-positive breast cancer with acquired resistance.
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