Lung Cancer Xenograft

Altogen Labs validated Lung Cancer Xenograft animal models:

A549, H460, H226, NCI-H1975, DMS273, LL/2, Calu-3, Calu-6, H1155, NCI-H522

Xenotransplantation studies have been a backbone of oncology research for four decades, and provide an effective research and evaluation environment for novel pharmaceutical compounds. Typically, these studies involve the implantation of tumorigenic human cell lines into immunocompromised mice, providing scientists with an in vivo model of tumor behavior in which to perform experiments including screening of novel cancer therapies, studies of cell behavior, and examination of metastasis. Patient-derived xenografts are a fundamental part of in vivo pharmacological research, aiding in the translation from benchtop to bedside.

Lung cancer that starts in the lung tissue is known as primary lung carcinoma, which are made of cancerous epithelial cells. Symptoms of lung cancer include excessive coughing, coughing blood, shortness of breath, weight loss and chest pains. There are risk factors of developing lung cancer that include smoking, genetics, asbestos, radon gas and air pollution. There are two primary types of lung cancer. The first is small cell lung carcinoma (SCLC) and the second is non-small-cell lung carcinoma (NSCLC). SCLC cells are characterized by the vesicles with neuroendocrine hormones carried by dense neurosecretory granules. SCLC is typically found in larger airways such as primary and secondary bronchi, and this cancer type presents as extensive and aggressive. NSCLC often has mutated epidermal growth factor receptor (EGFR) and dysregulated oncogenes including c-MET, LKB1, NKX2-1, BRAF and PIK3CA. The main substypes of NSCLC include adenocarcinoma, large-cell carcinoma and squamous cell carcinoma. As primary lung carcinoma are epithelial cells, metastasis requires epithelial to mesenchymal transition (EMT) which is aided through deregulation of signaling pathways including Fas, Par6, MEK-ERK and Akt/GSK3β. K-ras is also a commonly mutated proto-onocogene in lung adenocarcinomas. The lungs are also a common organ metastasis site for other types of primary cancers such as breast cancer. Diagnosis is typically with chest radiographs or CT scans, and a bronchoscopy biopsy can help with a more detailed evaluation. Prognosis for lung cancer in the US is poor, a 17.4% 5-year survival rate. Treatment of lung cancer includes minimizing exposure to risk factors such as smoking, surgery (needle aspiration, endoscopic ultrasound with needle aspiration, video-assistid thoracoscopic surgery (VATS), lobectomy, wedge resection, pneumonectomy,  and mediastinoscopy), radiotherapy (brachytherapy, external beam radiotherapy), chemotherapy (different for NSCLC and SCLC but may include carboplatin, gemcitabine, paclitaxel, cisplatin, irinotecan, pemetrexed, decetaxel) and targeted therapy (erlotinib, gefitinib, EGFR inhibitors, monoclonal antibodies against receptor activator of NF-kappa-B). Even when diagnosed at the earliest clinical stage IA, the 5-year survival rate is 50% or less.

Using human xenograft models of leukemia, as previously mentioned, is a powerful research tool, and there are several models of leukemia to choose from. There are links above to some of the most common tissue culture models that Altogen Labs has available, summarized in the table below. Models are often selected based on morphology, genetics, histology, early vs. late stage phenotype, invasive/aggressive properties, and abnormal protein expressions (usually relating to cell cycle, apoptosis, growth and angiogenesis). The goal of xenografts and murine models is to mirror human pathology and disease as closely as possible so that accurate insights into cellular events are achieved. This aspect is particularly critical with preclinical drug testing for accurately evaluating compound efficacy.

Cell Line	Characteristics
A549	·    Epithelial human cells from a skin metastasis site of a patient with pulmonary adenocarcinoma·    Hypotriploid karyotype·    Common model for allergies, respiratory infections and asthma ·    Exhibits ability to spontaneously metastasize in xenografts ·    NSCLC
H460	·    Epithelial human line from a patient with large-cell NSCLC·    Expresses p53, vimentin and keratin·    Does not express neurofilament triplet protein ·    No structural DNA abnormalities
H226	·    Derived from the pleural effusion of a patient diagnosed with mesothelioma.·    Squamous cell carcinoma (epithelial)·    Used in chemosensitivity testing
NCI-H1975	·    Epithelial NSCLC from a non-smoker patient with lung adenocarcinoma
LL/2	·    Established from a mouse that developed Lewis lung carcinoma·    Hypotetraploid karyotype·    Tumorigenic but weakly metastatic
Calu-3	·    Derived from the primary tissue of a patient with NSCLC adenocarcinoma·    Expresses HER2 and EGFR·    Differentiation is affected by cell culture conditions
Calu-6	·    Epithelial human cell line derived from a patient with pulmonary anaplastic carcinoma·    Hypotriploid karyotype·    Expresses renin, cytokeratin-7, cytokeratin-8, cytokeratin-18, cytokeratin-19 and vimentin
H1155	·    Epithelial cells derived from a metastasis in the lymph node originating from lung carcinoma prior to treatment·    Expresses EGFR and PD-L1·    NSCLC ·    Does not express gastrin releasing peptide (GRP)

DMS273

Human female lung small cell carcinoma from a pleural effusion Patient underwent therapy with cytoxan, methotrexate, vincristine, and radiation prior to harvesting Expresses retinoblastoma protein and mRNA Expresses antigens Leu 7 and My23 Expresses genes adrenocorticotropin, bombesin, corticotropin, calcitonin, beta endorphin, lipotropin, 17beta estradiol, parathormone, SRIF and OT-NP Positive for oncogenes c-myc, n-myc, c-raf, H-ras, K-ras, N-ras; negative for oncogenes v-fes, and v-fms

NCIH522

●        Human NSCLC from a smoker prior to treatment●        Hypotriploid karyotype ●        Codon 191 of the TP53 gene is mutated ●        Do not synthesize gastrin releasing peptide (GRP), neurofilament triplet protein or peptide neuromedin B (NMB) ●        Express keratin and vimentin

Altogen Labs is one of the leading biology contract research organization (CRO) based in Austin, Texas. Altogen Labs provides years of expert research in xenograft experiments taking advantage of the comprehensive expertise the company has developed in the use of human tumor xenografts for research and clinical purposes. Altogen Labs offers a complete suite of laboratory services, including:

• xenotransplantation study design
• selection of appropriate cancer model/cell line
• host animal selection
• subcutaneous or orthotopic xenografting
• daily observation of experimental subjects
• post-experiment analysis, including serum collection and histology

Mouse strains available at Altogen Labs:

Mouse type	T cells	B cells	NK cells	Coat	Other Notes
CD1	No	Yes	Yes	White/albino	Outbred
B6	Yes	Yes	Yes	White/albino	Inbred
Balb/c	No	Yes	Yes	Nude, albino	Inbred
Balb/SCID	No	No	Yes	White	Inbred
NOD/SCID	No	No	Impaired	White	Inbred
Nu/nu	No	Yes	Yes	Nude	Outbred
CD57BL	Yes	Yes	Yes	Dark brown/black	Inbred
CB17	Yes	Yes	Yes	White	Inbred
NSG	No	No	No	White	Inbred
Swiss Nude	No	Yes	Yes	Nude	Outbred

About the models

CD1

This model originates from a non-inbred Swiss stock of the 1920s from the Centre Anticancerux Romand (Lausanne, Switzerland). Outbred stocks are generally used for their genetic variability.

B6

This strain of mouse arose from a spontaneous mutation in the C57BL/6 strain resulting in a coisogenic albino mutant. These mice have a mutant tyrosinase gene.

Balb/c

This strain of nude mouse was developed in the 1980s through many crosses and backcrosses and remains to be an inbred model. Balb/c mice do not have a thymus and therefore cannot produce T-cells and are considered immunodeficient. Balb/c mice are often used for their easy breeding and similar weights (low-variation) of males and females. They are also used for monoclonal antibody production.

Balb/SCID

This mouse model lacks functioning T and B cells but do have functioning NK cells which limits engraftment. These mice are sensitive to irradiation and have functioning macrophages, dendritic cells and complement activity. Some cancer cell lines show improved engraftment over nude models in Balb/SCID mice.

NOD/SCID

The homozygous SCID mutation results in impaired T cell and B cell lymphocyte development. The NOD characteristic results in impaired natural killer cell function. NOD/SCID mice also lack macrophage and dendritic cell activity as well as reduced complement activity. These mice have a non-obese diabetic and insulitis background and low cytokine production. NOD/SCID mice exhibit a 36-week median survival due to the development of thymic lymphomas, which limits their use to short-term experiments.

Nu/nu

These mice originate from the National Institute of Health (NIH). Originally thought to be BALB/C congenic mice, once it was discovered that these mice were outbred they were determined to be of their own strain. These mice do not have a thymus, or T-cells, and are nude immunodeficient models.

CD57BL/6

This laboratory mouse strain was the 2^nd mammalian species to ever have its genome published in entirety. They originate from the Bussey Institute for Research in Applied Biology in 1921. These mice are often selected for easy breeding and availability of congenic strains. These mice are particularly sensitive to odors, noise, pain, cold, alcohol and morphine addiction.

CB17

CB17 mice are of a congenic strain that carry the immunoglobulin heavy chain allele (Igh-1b) from a C57BL/Ka on a BALB/c background. They are an ideal control for the CB17/SCID immunodeficient mouse model

NSG

Also known as NOD scid gamma, these mice are deficient in NK, T and B cells as well as multiple cytokine pathways. They also have reduced dendritic cell function and defective macrophage activity and lack a complement system. They are one of the most immunodeficient models available and unlike NOD/SCID mice, NSG mice do not develop thymic lymphomas and can be used for long-term experiments.

Swiss Nude

These mice originate from the 1974 Gustave Roussy Institute (Villejuif, France) Swiss stock. They are T cell deficient, nude and albino.

All laboratory studies are performed by experienced personnel in a GLP-compliant and IACUC-regulated facility in Austin, Texas. Please contact us at info@altogenlabs.com, or call 512-433-6177 to discuss xenograft study details.