PCET Expertise and
Resources that Drive Outcomes
Aragen’s highly experienced team will guide you through Pre-Clinical Efficacy Testing (PCET) by recommending the most appropriate animal and cell-based models, conducting studies with protocols and reports customized to your needs.
We offer a diverse range of oncology disease models and services to provide the most appropriate study design for the specific project needs. These models range from human xenograft tumor models (CDX) to the more complex sub-renal capsule, patient derived xenograft (PDX), and both syngeneic and humanized immuno-oncology models. Aragen’s services leverage our cutting edge technology platform and laboratories which include an 8,500 square-foot AAALAC accredited animal facility.
Human Cell Line Xenograft Models
There are hundreds of cell lines, various mouse strains and multiple implantation techniques that can be utilized in a xenograft model. Aragen has experience with xenograft models of lymphoma, leukemia, lung cancer, breast cancer, colon cancer, melanoma, and other indications challenged with both large- and small molecule therapeutics. These can be run in nude or other immunodeficient murine strains. Depending on the needs of the study and the mechanism of action of the molecules, these tumors can be implanted subcutaneously, orthotopically, within the sub-renal capsule or, in some cases, run as a metastatic model. We are also well trained on the various dosing methods and regiments required for these studies.
The most frequently and cost effectively used for the evaluation of in vivo efficacy of novel anticancer drug candidates. Tumor cells are subcutaneously implanted on the flank of animals and tumor is measured using vernier caliper.
In the orthotopic model, cells are introduced directly into their organ/tissue of origin. Aragen provides several orthotopic models including breast cancer, pancreatic, liver, and disseminated models. Other models may be developed upon request. When combined with the IVIS system, , insertion of a luciferase (bioluminescence) gene into the genome of the cell line of interest can visually track tumor growth in real-time throughout the in-life part of the study.
Representative whole body bioluminescence images following surgically implanted Bx-PC3 in athymic mice.
Patient Derived Xenograft (PDX) Tumor Models
Efforts have been made to create oncology models that more closely represent the biology and therapeutic responses observed in human patients. To that end, models have been generated in which tumor tissue derived directly from a human patient is implanted and grown in a immuno-compromised mouse. Aragen has experience with a number of commercially available PDX models.
Immuno-Oncology (Cancer Immunotherapy Models)
One of the most promising new areas of therapeutic development is immuno-oncology (I/O), which relies on triggering the body’s own immune system to fight cancer. Aragen offers a comprehensive selection of I/O preclinical models designed to fulfill a wide range of research tasks and efficacy testing. Since I/O seeks to activate the body’s natural immune response to fight cancer, it is fundamentally different than most treatment approaches and requires unique considerations in development, such as resistance, endpoint considerations and adverse reactions.
Adoptive Transfer of Human PBMCs or T Cells
With the adoptive hPBMCs transfer, or more specifically T cell transfer to mice, the human immune system can be partially reconstituted in these murine models. These “humanized mice” aim at harboring an immune environment capable of more accurately reflecting the tumor growth in the presence of I/O drugs. The injection route can be subcutaneous (where the PBMC are admixed with tumor cells), intravenously, or intraperitoneally. Tumor burden in the orthotopic or disseminated models can be routinely monitored by the IVIS system. Partial immune reconstitution during the efficacy testing period can be validated using FACS analysis for detection of human lymphocytes (CD45+CD3+) including other immune subpopulations (e.g. CD4+, CD8+). Aragen can assay your specific target molecule expressed in the xenograft prior to the tumor implantation with an antibody binding assay.
CAR-T Cell Therapy Efficacy Testing
Immunotherapy using T cells genetically engineered to express a chimeric antigen receptor (CAR) is emerging as a promising new treatment for certain lymphomas and leukemias. Preclinical assessment of CAR-T-cell efficacy and safety are necessary for dosage determination and risk management. CAR-T-cell therapy can induce a rapid and sustained clinical response, which are sometimes associated with acute toxicities. Our efficacy testing includes the establishment of the xenograft model, CAR-T administration, and efficacy evaluation. Evaluation of CAR-T induced toxicity (e.g. cytokine analysis), as well as animal necropsy for endpoint evaluation with sample collection (e.g. serum, tumor at the endpoint) can be additionally included. The activity of transferred CAR-T can be routinely monitored by the IVIS system or FACS analysis.
If the mechanism of action requires an intact immune system, the immuno-compromised setting required for a xenograft model will not be appropriate for efficacy assessment. In such cases syngeneic models, where the strain of the derived tumor cell line and the host animal are the same, may be more appropriate. Aragen has performed many studies using syngeneic models including models of breast, colon, liver, and lung cancers as well as melanoma and lymphoma models.
Syngeneic mouse models can provide a comprehensive, effective approach to help understand the underlying mechanism(s) of candidate immune-therapeutics in the presence of a functionally immunocompetent system. with historical cell line growth and drug sensitivity data. Aragen has performed many syngeneic studies including models of breast, colon, liver, and lung cancers as well as models of melanoma and lymphoma. We can perform complex efficacy regimen including combination drug studies with various routes of administration. Immunophenotyping by flow cytometry enables identification of models that harbor “hot (inflamed)” or “cold (non-inflamed)” tumor immune infiltrate phenotypes. These responses, along with cross talk between tumor and immune system in the tumor microenronment, are translatable to the patient responses in clinical settings.
Immune Response Profiling
We leverage our broad experience and state-of-the-art technologies to analyze tumor-infiltrating lymphocytes (TIL), markers in the tumor microenvironment (TME), and in peripheral sites (eg. blood, spleen, lymph nodes) using appropriate quantitative multiplex technologies such as flow cytometry, IDEXX ProCyte DX Hematology Analyzer, RT-qPCR, and multiplex IQ analyzer.
CBC analysis can be performed separately or in combination with other immuno-assays as a complementary immuno-assay to assess disease progression. For example, myelosuppression induced by conventional “standard-care” therapeutics (commonly known as chemotherapeutics) can be detected immediately from a small blood draw. Alternatively, certain inflammatory response profiles can be correlated with tumor burden. Drug or cytokine-induced lymphocyte proliferation, or drug induced toxicities such as anemia or changes in platelet counts can readily identified, making them an ideal companion diagnostic tool for critical therapeutic decisions.
Multiplex Immunophenotyping Capability
Our 14-color FACS analysis platform allow us to evaluate both human and murine immunophenotyping subsets by selectively gating immune cells of interest.
Analysis of Effector and Memory Markers in CD8+ T Cell Subsets in Blood Sample from CT26 Bearing Mouse (Left) or Tumor Free Survivor (Right).
Other comprehensively customized pre-clinical immuno-oncology assays includes:
- Immune Cell Activation/Proliferation
- MLR Assay
- CTL Assay
- ADCC Assay
- ADCP Assay
- Phagocytosis/ ROS Assay
Our broad in vivo services expertise enables flexibility in the way we utilize our technology platforms. This allows us to tailor our methods to meet your project requirements for quality and assay performance.
- PerkinElmer IVIS XRMS Imaging System
- ThermoFisher Attune Flow Cytometer (14 color)
- Flexivent Lung Function Analyzer
- Whole Body Plethysmograph
- IDEXX ProCyte Dx Hematology Analyzer
- Catalyst Dx Chemistry Analyzer
- Multimodal Plate Reader
- Fluorescent Microscope
- Standard Cell Culture
- BSL-2 Containment
- Microsurgical Instrumentation
- Cell Culture
- Micro-scale Doppler Probes
- IntelliCyt IQue Screener