University of Texas MD Anderson Cancer Center 17 articles published in JoVE Cancer Research Generation and Expansion of Primary, Malignant Pleural Mesothelioma Tumor Lines Tamara M. Griffiths1, Carlos Ramos1, Cara L. Haymaker1 1Department of Translational Molecular Pathology, University of Texas MD Anderson Cancer Center The goal of this method paper is to demonstrate a robust and reproducible methodology for the enrichment, generation, and expansion of primary tumor cell lines from surgically resected pleural mesothelioma. Neuroscience Mechanical Conflict-Avoidance Assay to Measure Pain Behavior in Mice Caitlyn M. Gaffney1, Gabriella Muwanga2, Huaishuang Shen2, Vivianne L. Tawfik2, Andrew J. Shepherd2 1Laboratories of Neuroimmunology, Department of Symptom Research, and the MD Anderson Pain Research Consortium, University of Texas MD Anderson Cancer Center, 2Department of Anesthesiology, Perioperative and Pain Medicine, Stanford University School of Medicine The mechanical conflict-avoidance assay is used as a non-reflexive readout of pain sensitivity in mice which can be used to better understand affective-motivational responses in a variety of mouse pain models. Biology Visualization of DNA Repair Proteins Interaction by Immunofluorescence Bárbara de la Peña Avalos1,2, Eloïse Dray1,2 1Department of Biochemistry and Structural Biology, University of Texas Health Science Center at San Antonio, 2Mays Cancer Center at UT Health San Antonio MD Anderson Cancer Center Following DNA damage, human cells activate essential repair pathways to restore the integrity of their genome. Here, we describe the method of indirect immunofluorescence as a means to detect DNA repair proteins, analyze their spatial and temporal recruitment, and help interrogate protein-protein interaction at the sites of DNA damage. Medicine Radiation Planning Assistant - A Streamlined, Fully Automated Radiotherapy Treatment Planning System Laurence E. Court1, Kelly Kisling1, Rachel McCarroll1, Lifei Zhang1, Jinzhong Yang1, Hannah Simonds2, Monique du Toit2, Chris Trauernicht2, Hester Burger3, Jeannette Parkes3, Mike Mejia4, Maureen Bojador4, Peter Balter1, Daniela Branco1, Angela Steinmann1, Garrett Baltz1, Skylar Gay1, Brian Anderson1, Carlos Cardenas1, Anuja Jhingran5, Simona Shaitelman5, Oliver Bogler6, Kathleen Schmeller7, David Followill1, Rebecca Howell1, Christopher Nelson1, Christine Peterson8, Beth Beadle5,9 1Department of Radiation Physics, University of Texas MD Anderson Cancer Center, 2Department of Radiation Oncology, Stellenbosch University and Tygerberg Hospital, 3Departments of Radiation Oncology and Medical Physics, Groote Schuur Hospital and University of Cape Town, 4Department of Radiation Oncology, University of Santo Tomas Hospital, Benavides Cancer Institute, 5Department of Radiation Oncology, University of Texas MD Anderson Cancer Center, 6Academic Affairs, University of Texas MD Anderson Cancer Center, 7Department of Gynecological Oncology and Reproductive Medicine, University of Texas MD Anderson Cancer Center, 8Department of Biostatistics, University of Texas MD Anderson Cancer Center, 9Department of Radiation Oncology, Stanford University Radiation therapy is a highly complex cancer treatment that requires multiple specialists to create a treatment plan and provide quality assurance (QA) prior to delivery to a patient. This protocol describes the use of a fully automated system, the Radiation Planning Assistant (RPA), to create high-quality radiation treatment plans. Cancer Research An Integrated Platform for Genome-wide Mapping of Chromatin States Using High-throughput ChIP-sequencing in Tumor Tissues Christopher Terranova1, Ming Tang1, Elias Orouji1, Mayinuer Maitituoheti1, Ayush Raman1, Samirkumar Amin2, Zhiyi Liu1, Kunal Rai1 1Department of Genomic Medicine, University of Texas MD Anderson Cancer Center, 2The Jackson Laboratory for Genomic Medicine Here, we describe an optimized high-throughput ChIP-sequencing protocol and computational analyses pipeline for the determination of genome-wide chromatin state patterns from frozen tumor tissues and cell lines. Immunology and Infection In Vivo Assay for Detection of Antigen-specific T-cell Cytolytic Function Using a Vaccination Model Cara L. Haymaker*1, Yared Hailemichael*1, Yi Yang2,3, Roza Nurieva2 1Department of Melanoma Medical Oncology, University of Texas MD Anderson Cancer Center, 2Department of Immunology, University of Texas MD Anderson Cancer Center, 3Department of Radiation Oncology, The Second Hospital of Jilin University The goal of this protocol is to allow for detection of in vivo antigen-specific killing of a target cell in a murine model. Cancer Research Preparation of Primary Acute Lymphoblastic Leukemia Cells in Different Cell Cycle Phases by Centrifugal Elutriation Magdalena Delgado*1, Anisha Kothari*1, Walter N. Hittelman2, Timothy C. Chambers1 1Department of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences, 2Department of Experimental Therapeutics, University of Texas MD Anderson Cancer Center This protocol describes the use of centrifugal elutriation to separate primary acute lymphoblastic leukemia cells into different cell cycle phases. Genetics Analysis of the c-KIT Ligand Promoter Using Chromatin Immunoprecipitation Pingyu Zhang*1, Andres Rojas*1, Boris Blechacz1 1Department of Gastroenterology, Hepatology and Nutrition, The University of Texas MD Anderson Cancer Center DNA-protein interactions are essential for multiple biological processes. During the evaluation of cellular functions, the analysis of DNA-protein interactions is indispensable for understanding gene regulation. Chromatin immunoprecipitation (ChIP) is a powerful tool to analyze such interactions in vivo. Biology En Face Preparation of Mouse Blood Vessels Kyung Ae Ko1, Keigi Fujiwara1, Sunil Krishnan2, Jun-ichi Abe1 1Department of Cardiology, Division of Internal Medicine, University of Texas MD Anderson Cancer Center, 2Department of Radiation Oncology, University of Texas MD Anderson Cancer Center A procedure for making en face preparations of the mouse carotid artery and aorta is described. Such preparations, when immunofluorescently stained with specific antibodies, enable us to study localization of proteins and identification of cell types within the entire vascular wall by confocal microscopy. Biology Sample Preparation for Mass Cytometry Analysis Ryan L. McCarthy1, Aundrietta D. Duncan1, Michelle C. Barton1 1Epigenetics and Molecular Carcinogenesis, The University of Texas MD Anderson Cancer Center This article describes the collection and processing of samples for mass cytometry analysis. Neuroscience Novel Assay for Cold Nociception in Drosophila Larvae Heather N. Turner1,2,5, Christian Landry3, Michael J. Galko1,2,4 1Department of Genetics, UT MD Anderson Cancer Center, 2Neuroscience Program, Graduate School of Biomedical Sciences at Houston, 3ProDev Engineering, 4Genes and Development Program, Graduate School of Biomedical Sciences at Houston, 5Section of Neurobiology, University of Southern California Here we demonstrate a novel assay to study cold nociception in Drosophila larvae. This assay utilizes a custom-built Peltier probe capable of applying a focal noxious cold stimulus and results in quantifiable cold-specific behaviors. This technique will allow further cellular and molecular dissection of cold nociception. Cancer Research Intracarotid Cancer Cell Injection to Produce Mouse Models of Brain Metastasis Chenyu Zhang1, Frank J. Lowery1, Dihua Yu1 1Department of Molecular and Cellular Oncology, The University of Texas MD Anderson Cancer Center Brain metastasis has become an urgent unmet medical need as its incidence has increased while therapeutic options have remained palliative. Creating experimental animal models of brain metastasis via intracarotid arterial injection of cancer cells facilitates mechanistic studies of the disease biology and evaluation of novel intervention regimens. Developmental Biology Technique to Target Microinjection to the Developing Xenopus Kidney Bridget D. DeLay1, Vanja Krneta-Stankic1,2, Rachel K. Miller1,2,3,4 1Department of Pediatrics, Pediatric Research Center, University of Texas McGovern Medical School, 2Program in Genes & Development, University of Texas Graduate School of Biomedical Sciences, 3Program in Cell & Regulatory Biology, University of Texas Graduate School of Biomedical Sciences, 4Department of Genetics, University of Texas MD Anderson Cancer Center Here, we present a protocol to use fate maps and lineage tracers to target injections into individual blastomeres that give rise to the kidney of Xenopus laevis embryos. Medicine Working with Human Tissues for Translational Cancer Research Alexandre Reuben*1, Vancheswaran Gopalakrishnan*1, Heidi E. Wagner3, Christine N. Spencer2, Jacob Austin-Breneman1, Hong Jiang1, Zachary A. Cooper1, Jennifer A. Wargo1 1Department of Surgical Oncology, University of Texas MD Anderson Cancer Center, 2Department of Genomic Medicine, University of Texas MD Anderson Cancer Center, 3Department of Pathology and Institutional Tissue Bank, University of Texas MD Anderson Cancer Center Translational cancer research is dependent on extraction of human tissues. Much work has gone into optimizing extraction methods for ex vivo analysis. Here, we describe tissue processing methods allowing for maximal data output from limited samples. Immunology and Infection Automated Cell Enrichment of Cytomegalovirus-specific T cells for Clinical Applications using the Cytokine-capture System Pappanaicken Kumaresan*1, Mathew Figliola*1, Judy S. Moyes1, M. Helen Huls1, Priti Tewari1, Elizabeth J. Shpall2, Richard Champlin2, Laurence J.N. Cooper1 1Division of Pediatrics, U.T. MD Anderson Cancer Center, 2Stem Cell Transplantation and Cellular Therapy, U.T. MD Anderson Cancer Center The goal of this protocol is to manufacture pathogen-specific clinical-grade T cells using a bench-top, automated, second generation cell enrichment device that incorporates a closed cytokine capture system and does not require dedicated staff or use of a GMP facility. The cytomegalovirus pp65-specific-T cells generated can be directly administered to patients. Medicine Utilizing Murine Inducible Telomerase Alleles in the Studies of Tissue Degeneration/Regeneration and Cancer Takashi Shingu1, Mariela Jaskelioff2, Liang Yuan1, Zhihu Ding3, Alexei Protopopov4, Maria Kost-Alimova4, Jian Hu1 1Department of Cancer Biology, UT MD Anderson Cancer Center, 2Novartis Institutes for Biomedical Research, 3Sanofi US, 4Institute of Applied Cancer Science, UT MD Anderson Cancer Center Telomere and telomerase play essential roles in ageing and tumorigenesis. The goal of this protocol is to show how to generate two murine inducible telomerase knock-in alleles and how to utilize them in the studies of tissue degeneration/regeneration and cancer. Medicine Quantitative Multispectral Analysis Following Fluorescent Tissue Transplant for Visualization of Cell Origins, Types, and Interactions Erika L. Spaeth1, Christopher M. Booth2, Frank C. Marini1,2 1Department of Leukemia, MD Anderson Cancer Center, 2Wake Forest Baptist Medical Center, Institute for Regenerative Medicine Complex tissue masses, from organs to tumors, are composed of various cellular elements. We elucidated the contribution of cellular phenotypes within a tissue utilizing multi-labeled fluorescent transgenic mice in combination with multiparameter immunofluorescent staining followed by spectral unmixing to decipher cell origin as well as cell characteristics based on protein expression.