Lawrence W. Anderson

New cell line. Epithelial cell line and subline established from premalignant mouse mammary tissue.

SummaryA cell line and subline with epithelial characteristics were established from mouse mammary hyperplastic alveolar nodules (HAN). The cells do not grow in suspension cultures in vitro or form tumors in vivo. The cells do produce significant amounts of C-type and A-type virus and low amounts of plasminogen activator.

Metastatic Potential of Hyperplastic Alveolar Nodule Derived Mouse Mammary Tumor Cells Following Intravenous Inoculation

Abstract A comparison was made of the in vivo growth potential of tumor cells derived from mouse mammary adenocarcinomas arising in premalignant outgrowth lines D1 and D2. Twenty-one tumors were dissociated and cultured as cell monolayers in vitro. Three to four days later 50,000–100,000 cells were inoculated intravenously into syngeneic female BALBlc mice. Primary cultures of tumor cells which arose in D2 HAN outgrowths varied considerably in their ability to form tumor nodules in the lungs of syngeneic mice 4 weeks post-inoculation. An increase in either the number of cells inoculated or in the length of in vivo incubation influenced the extent of tumor growth observed in the lungs. Primary cultures of tumor cells which arose in D1 HAN outgrowths inoculated at the same cell number rarely formed tumor nodules in the lungs 4 weeks post-injection. If, however, twice as many cells were introduced, a small number of large nodules were observed within 4 weeks. An epithelial cell line (WAZ-2T) isolated from D1 tumor tissue produced small lung nodules by 4 weeks, but larger more numerous nodules arose when in vivo incubation was extended an additional 3 weeks. This study indicates that primary D2 tumor cells have a variable but higher degree of metastatic potential than primary D1 tumor cells following intravenous inoculation. Tumors which develop in the D2 and D1 premalignant lines may be useful for studies of chemotherapy and cell latency of tumor metastases and the identification of the specific cell subpopulations involved in these events.

The National Cancer Institute ALMANAC: A Comprehensive Screening Resource for the Detection of Anticancer Drug Pairs with Enhanced Therapeutic Activity

To date, over 100 small-molecule oncology drugs have been approved by the FDA. Because of the inherent heterogeneity of tumors, these small molecules are often administered in combination to prevent emergence of resistant cell subpopulations. Therefore, new combination strategies to overcome drug resistance in patients with advanced cancer are needed. In this study, we performed a systematic evaluation of the therapeutic activity of over 5,000 pairs of FDA-approved cancer drugs against a panel of 60 well-characterized human tumor cell lines (NCI-60) to uncover combinations with greater than additive growth-inhibitory activity. Screening results were compiled into a database, termed the NCI-ALMANAC (A Large Matrix of Anti-Neoplastic Agent Combinations), publicly available at https://dtp.cancer.gov/ncialmanac Subsequent in vivo experiments in mouse xenograft models of human cancer confirmed combinations with greater than single-agent efficacy. Concomitant detection of mechanistic biomarkers for these combinations in vivo supported the initiation of two phase I clinical trials at the NCI to evaluate clofarabine with bortezomib and nilotinib with paclitaxel in patients with advanced cancer. Consequently, the hypothesis-generating NCI-ALMANAC web-based resource has demonstrated value in identifying promising combinations of approved drugs with potent anticancer activity for further mechanistic study and translation to clinical trials. Cancer Res; 77(13); 3564-76. ©2017 AACR.

An in vitro model of neoplastic progression in murine epithelial cells

We have developed an epithelial cell line derived from mouse liver which represents a spectrum of malignant progression. When inoculated into mice at low passage, the cells induced benign cysts; after extensive subcultures, the same line induced low grade adenocarcinomas. A variant of these cells with increased invasive and metastatic potential was selected. In culture, growth in methocel correlated with acquisition of malignant potential while increased homotypic aggregation correlated with metastatic ability. These cultures should be extremely valuable for studies on the nature of epithelial cell malignancy, the most common form of human cancer.

Abstract 2306: Pre-clinical development of 4′-thio-2′-deoxycytidine (TdCyd) as a DNA-demethylating agent for use in treating solid tissue tumors

Targeting cancer epigenetic control of cell growth via DNA methylation has been successful in treating hematologic diseases, such as Decitabine (DAC) and Azacitidine for Myelodysplastic Syndrome including Acute Myelomonocytic Leukemia. This success has not extended to solid tissue tumors. The Division of Cancer Treatment and Diagnositcs of NCI has initiated pre-clinical development of TdCyd as an agent for treating solid tumors after promising early results in a lung adenocarcinoma xenograft model (NCI-H23). IP dosing at 5MKG (0.56 MTD) in nu/nu mice on a Q5D x 3 cycle schedule resulted in tumor stasis with no accompanying weight loss in the mice. A DAC-treated control arm treated at MTD resulted in tumor growth delay but not stasis, and a 10% weight loss was noted. Intratumoral levels of DNMT1 were reduced to undetectable levels in xenografts post administration of TdCyd by ELISA and Western Blot assays, but were unaffected by DAC treatment. Mass Spectrometry analysis demonstrated incorporation of both TdCyd and thiothymidine (TdThd) into H23 DNA. In vitro experiments on a selected panel of cancer cell lines confirmed the conversion of TdCyd to the triphosphate and re-expression of tumor suppressor proteins p15 and p16. Funded by NCI Contract No. HHSN261200800001E. Citation Format: Robert J. Kinders, Melinda Hollingshead, Jaideep Thottassery, William B. Parker, Thomas D. Pfister, Lawrence W. Anderson, Joseph E. Tomaszewski, Jerry M. Collins, James H. Doroshow. Pre-clinical development of 4′-thio-2′-deoxycytidine (TdCyd) as a DNA-demethylating agent for use in treating solid tissue tumors. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2306. doi:10.1158/1538-7445.AM2014-2306

Simian Virus 40 and Moloney-Murine Sarcoma Virus Infection of Bona fide Mouse Epithelium

Moloney-murine sarcoma virus (Moloney-MSV) and simian virus 40 (SV40) were found to infect successfully pure cultures of epithelial cells established from mouse liver and mammary tissue. MSV infection resulted in transient morphological foci with persistent production of infectious virus. SV40 infection produced detectable levels of virus-specific T antigen in the cells but morphological transformants were not observed.