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1. Discovery-based science education: functional genomic dissection in Drosophila by undergraduate researchers.

Author

Jiong Chen, Gerald B Call, Elsa Beyer, Chris Bui, Albert Cespedes, Amy Chan, Jenny Chan, Stacy Chan, Akanksha Chhabra, Peter Dang, Artemis Deravanesian, Brenda Hermogeno, James Jen, Eunha Kim, Eric Lee, Gemma Lewis, Jamie Marshall, Kirsten Regalia, Farnaz Shadpour, Aram Shemmassian, Kristin Spivey, Maggie Wells, Joy Wu, Yuki Yamauchi, Amir Yavari, Anna Abrams, Amanda Abramson, Latiffe Amado, Jenny Anderson, Keenan Bashour, Elena Bibikova, Allen Bookatz, Sarah Brewer, Natalie Buu, Stephanie Calvillo, Joseph Cao, Aileen Chang, Daniel Chang, Yuli Chang, Yibing Chen, Joo Choi, Jeyling Chou, Sumit Datta, Ardy Davarifar, Poonam Desai, Jordan Fabrikant, Shahbaz Farnad, Katherine Fu, Eddie Garcia, Nick Garrone, Srpouhi Gasparyan, Phyllis Gayda, Chad Goffstein, Courtney Gonzalez, Mariam Guirguis, Ryan Hassid, Aria Hong, Julie Hong, Lindsay Hovestreydt, Charles Hu, Farid Jamshidian, Katrin Kahen, Linda Kao, Melissa Kelley, Thomas Kho, Sarah Kim, Yein Kim, Brian Kirkpatrick, Emil Kohan, Robert Kwak, Adam Langenbacher, Santino Laxamana, Chris Lee, Janet Lee, So-Youn Lee, To Hang S Lee, Toni Lee, Sheila Lezcano, Henry Lin, Peter Lin, Julie Luu, Thanh Luu, Will Marrs, Erin Marsh, Sarah Min, Tanya Minasian, Amit Misra, Miles Morimoto, Yasaman Moshfegh, Jessica Murray, Cynthia Nguyen, Kha Nguyen, Ernesto Nodado, Amanda O'Donahue, Ndidi Onugha, Nneka Orjiakor, Bhavin Padhiar, Mara Pavel-Dinu, Alex Pavlenko, Edwin Paz, Sarah Phaklides, Lephong Pham, Preethi Poulose, Russell Powell, Aya Pusic, Divi Ramola, Meghann Ribbens, Bassel Rifai, Desiree Rosselli, Manyak Saakyan, Pamela Saarikoski, Miriam Segura, Ramnik Singh, Vivek Singh, Emily Skinner, Daniel Solomin, Kosha Soneji, Erika Stageberg, Marina Stavchanskiy, Leena Tekchandani, Leo Thai, Jayantha Thiyanaratnam, Maurine Tong, Aneet Toor, Steve Tovar, Kelly Trangsrud, Wah-Yung Tsang, Marc Uemura, Mary Unkovic, Emily Vollmer, Emily Weiss, Damien Wood, Sophia Wu, Winston Wu, Qing Xu, Kevin Yackle, Will Yarosh, Laura Yee, George Yen, Grant Alkin, Sheryllene Go, Devon M Huff, Helena Minye, Eric Paul, Nikki Villarasa, Allison Milchanowski, and Utpal Banerjee

Subjects
Biology (General), QH301-705.5
Published
2005
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2. Genomewide Clonal Analysis of Lethal Mutations in the Drosophila melanogaster Eye: Comparison of the X Chromosome and Autosomes

Author

Christopher Lee, Thanh Luu, Maurine Tong, Cheryl Chan, Thomas Walsh, Mabel Chin, Jiong Chen, Jacob Makshanoff, Diane Vo, Yuli Chang, John M. Olson, Tony Hung, Yuki Yamauchi, Manyak Saakyan, Renata P. Lerner, Elizabeth Evers, Latiffe Amado, Nick Garrone, Carol Salame, Jesse M. Zaretsky, Eric Paul, Winston Wu, Trinh Tran, David Kohanchi, James Z. Hui, Neetika Srivastava, Allison M. Yabroff, Aria Hong, Daniel Solomin, Preethi Poulose, Brian Kirkpatrick, Katherine Wu, Eunha Kim, Abbas Anwar, Diane Tan, Nneka Orjiakor, Utpal Banerjee, Lindsay Hovestreydt, Riju Gupta, Michael Hsieh, Jian-ya Lin, Jennifer Leighn. Sta.Ana, Alex Pavlenko, Ardy Davarifar, Stephanie S. Latterman, Shekhar S. Pai, Qing Xu, Phyllis Gayda, Amir Yavari, Allen Bookatz, Kathy T. Ngo, Paul D. Picard, Srpouhi Gasparyan, ToHang S. Lee, Emily Skinner, Sanjit Datta, Jennifer D. Trinh, Sarah Brewer, Courtney Gonzalez, Matteo Pellegrini, Desiree Rosselli, Gerald B. Call, Gemma Lewis, Kosha Soneji, Quynh-Minh Tran, Amy W. Ku, Farzin Farshidi, Cynthia Nguyen, Vivek Singh, Tanya Minasian, Mara Pavel-Dinu, Chad Goffstein, Bryon C. Lim, So-Youn Lee, Stephanie S. Lauw, Long H. Nguyen, Kevin Yackle, Will Yarosh, Santino Laxamana, Chelsea E. Taber, Sarah Kim, Tiffany Liu, Stacy Chan, Armonde Baghdanian, Kelly Trangsrud, Robert Kwak, Gelsey L. Goodstein, Farnaz Shadpour, Mariam Guirguis, Wah-Yung Tsang, Jayantha Thiyanaratnam, Edwin Paz, Chloe Knudsen-Robbins, Tatiana Moroz, Arlene J. Formanes, Mei L. Iao, Latifat Alli, Shawn S. Badal, Kathleen Lin, PhuongThao Tran, Jamie L. Marshall, Regan S. Wong, Akanksha Chhabra, Ran X. Zhu, Amanda Abramson, Sarah Min, Amit Misra, Vanessa Soma, Blake E. Haas, Eddie Garcia, Ndidi Onugha, Divi Ramola, Ya-Whey Wu, Bogdana Kovshilovskaya, Crystal P. Y. Lui, Kai C. Lei, Jeyling Chou, Yasaman Moshfegh, Sohrab Sidhu, Mark E. L. Landig, Michael G. Yeranosian, James S. Yu, Laura Yee, Lydia S-H. Kim, Katrina Fischer, Angela Y. Kim, Junia J. Song, Melissa M. Kelley, Sheila Lezcano, Jingwen Tan, Vincent Luong, Jennifer J. Zhou, Christopher N.H. Bui, Jenny Chan, Sophia D. Jin, Natalie Buu, Emily Weiss, Amanda O'Donahue, Christian Sanchez, Andrew Folick, Nkau X. V. Yang, Christie Sun, Lephong Pham, Stephanie Calvillo, Charles Hu, Yein Kim, Aileen Chang, George Yen, Joy Wu, Poonam Desai, Linda Kao, Emily G. Travis, Mary Unkovic, Damien Wood, Miriam Segura, Marc Uemura, Henry Lin, Maggie Wells, Miles Morimoto, Sumit Datta, Kha Nguyen, Jessica Wickland, Christina C. Seifert, Leo Thai, Bhavin Padhiar, Hei J. Chan, Yibing Chen, Abhishek Shah, Julie Hong, Cheng R. Huang, Sara A. Baumann, Xibin A. Cheng, Russell Powell, Shawn J. Cokus, Sonia Arevalo, Anna L. Abrams, Mark Engeln, Minh-Tu Do, Joseph Cao, JiEun Lee, Brenda Hermogeno, Bassel Rifai, Preethika S Ekanayake, Alison S. Teruya, Carlyn Crisostomo, Lesley L. Leung, Toni Lee, Megan King, Amy Chan, William R. Marrs, Ryan Hassid, Elena Bibikova, Adam D. Langenbacher, Francie Diep, Jun Gong, Joann S. Lee, Manish Pandya, Emil Kohan, Daniel S. Lee, Jenny Anderson, Kristin Spivey, Devon M Huff, Yulee Kim, Thomas Kho, Alma Rangel, Janet Lee, Aneet Toor, Pamela Saarikoski, Nikki Villarasa, Julie Luu, Vivian N. Becerra, Yara A. Mikhaeil, Shahbaz Farnad, Vida Chitsazzadeh, Q. Angela Fang, Ju An, Leena Tekchandani, Tharani Prasad, Elsa Beyer, Christopher M. Allen, Fleurette Chong, Eric Lee, Meghann Ribbens, Sherrylene Go, Amrita K. Cheema, Jordan Fabrikant, Michael Safaee, An-Chi Mei, Peter Lin, Jessica Murray, Nina Sanchez, Katherine Fu, Sacha L. Prashad, Artemis Deravanesian, Erin Marsh, Robyn Tikia, Raymond W. Kung, Sarah Phaklides, Oganes H. Shilgevorkyan, Peter Dang, Aadit R. Patel, Erika Stageberg, Helena Minye, Alexander N. Patananan, Albert Cespedes, Kasey Topp, Vicky Hahm, Keenan Bashour, Joo Choi, Daniel Chang, Majid Moarefi, William Kim, Farid Jamshidian, Inderroop Singh, Aileen E. Chang, Miguel Meza, Edward Kuoy, Danny Ayoub, Aya Pusic, Daniel Sitz, Marina Stavchanskiy, Sophia Wu, Emily Vollmer, Angela Delosreyes, James Jen, Ernesto Nodado, Aram Shemmassian, Kirsten Regalia, Ramnik Singh, Arthur H. Baghdanian, Steve Tovar, H-C. Angela Liu, and Katrin Kahen

Subjects
Genetics Education, Genetics, Mutation, Genes, Essential, X Chromosome, Autosome, Genome, Insect, Genes, Insect, Biology, Eye, biology.organism_classification, medicine.disease_cause, Clonal analysis, Clone Cells, Drosophila melanogaster, medicine, Eye development, Animals, Genes, Lethal, Drosophila (subgenus), Gene, X chromosome
Abstract

Using a large consortium of undergraduate students in an organized program at the University of California, Los Angeles (UCLA), we have undertaken a functional genomic screen in the Drosophila eye. In addition to the educational value of discovery-based learning, this article presents the first comprehensive genomewide analysis of essential genes involved in eye development. The data reveal the surprising result that the X chromosome has almost twice the frequency of essential genes involved in eye development as that found on the autosomes.

Published
2007
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3. Functional Characterization of a T Cell Stimulation Reagent for the Production of Therapeutic Chimeric Antigen Receptor T Cells

Author

Valerie Odegard, Christopher Glen Ramsborg, Keenan Bashour, Ryan P Larson, Christian Stemberger, Patricia Graef, and Germeroth Lothar

Subjects
Chemistry, T cell, Immunology, T-cell receptor, CD28, Juno Therapeutics, Cell Biology, Hematology, Streptamer, Biochemistry, Chimeric antigen receptor, Cell biology, medicine.anatomical_structure, medicine, Cytotoxic T cell, IL-2 receptor
Abstract

Adoptive cell therapy using gene-modified T cells has demonstrated promising clinical outcomes in hematologic malignancies. Production of gene-modified T cells involves the selection of patient T cells, activation via stimulation through the endogenous T cell receptor (TCR) complex and a costimulatory domain, followed by introduction of a tumor antigen-specific TCR or chimeric antigen receptor (CAR) through gene modification. Here we characterize a soluble T cell stimulation reagent, known as an ExpamerTM reagent, in the production of therapeutic CAR T cells. The Expamer reagent used in these studies is designed to be a late-stage clinical and commercial manufacturing ancillary material with two important attributes that make it highly attractive from a manufacturing and regulatory standpoint; it is a soluble and dissociable reagent. These attributes increase the ease of both introduction and removal from the manufacturing process, giving products manufactured with this reagent consistent product quality and purity. This reagent activates T cells through the simultaneous engagement of the TCR-CD3 complex and the costimulatory receptor CD28 and is compatible with manufacturing of both current and next-generation therapeutics. Purified healthy donor T cells cultured in the presence of the Expamer reagent rapidly fluxed Ca2+, demonstrating the capacity to induce early TCR signaling. Activation through this reagent additionally promotes upregulation of the cell surface activation marker CD25 and proliferation as measured by CFSE dilution. Following stimulation with this reagent, T cells are readily transduced with a CD19-specific CAR construct. The function of CAR T cells generated with this reagent was measured by effector cytokine production, proliferation, and cytolytic activity in the presence of CD19 expressing and control target cells in vitro. CAR T cells robustly produced IFN-ɣ and IL-2 after activation with a CAR specific antigen. In addition, proliferation in the presence of CD19 expressing target cells was observed as measured by CFSE dilution. Finally, significant cytolytic activity against CD19-expressing target cells was observed. Collectively, these data provide evidence that functional engineered T cells can be manufactured using the Expamer reagent and support implementation into the production of both current and next-generation therapeutic gene-modified T cells. The first two authors contributed equally to this work. Disclosures Bashour: Juno Therapeutics: Employment. Larson:Juno Therapeutics: Employment. Graef:Juno Therapeutics: Employment. Stemberger:Juno Therapeutics: Employment. Lothar:Juno Therapeutics: Employment. Odegard:Juno Therapeutics: Employment. Ramsborg:Juno Therapeutics: Employment.

Published
2015
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4. Substrate rigidity regulates human T cell activation and proliferation (52.9)

Author

Roddy O'Connor, Xueli Hao, Keyue Shen, Keenan Bashour, Lance Kam, and Michael Milone

Subjects
Immunology, Immunology and Allergy
Abstract

Adoptive immunotherapy using cultured T cells holds promise for the treatment of cancer and infectious disease. Culture platforms based upon hard materials, such as polystyrene plastic, form the basis of many culture systems. The mechanical properties of a culture substrate can influence cellular adhesion, proliferation, and differentiation. We explored the impact of substrate stiffness on ex vivo T cell activation and polyclonal expansion using substrates with variable rigidity manufactured from poly(dimethylsiloxane) (PDMS), a biocompatible silicone elastomer. We show that the IL-2 production and ex vivo proliferation of human CD4+ and CD8+ T cells are increased an average of 4-fold following stimulation on softer (Young’s Modulus [E] < 100 kPa) compared with stiffer (E >2 MPa) substrates. Mixed peripheral blood T cells cultured on the stiffer substrates also demonstrate a trend towards a greater proportion of CD62Lneg, effector-differentiated CD4+ and CD8+ T cells. Culture of naïve CD4+ T cells on softer substrates yields an average 3-fold greater proportion of IFN-γ producing TH1-like cells. These findings reveal that the rigidity of the substrate used to immobilize T cell stimulatory ligands is an important and previously unrecognized parameter for T cell culture systems used for adoptive immunotherapy. These results also have implications for studies of T cell activation and signal transduction that use immobilized TCR/CD3 and CD28 ligands.

Published
2012
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5. Discovery-based science education: functional genomic dissection in Drosophila by undergraduate researchers

Author

Kristin Spivey, Aneet Toor, Gerald B. Call, Brenda Hermogeno, Pamela Saarikoski, Qing Xu, Sheryllene Go, Vivek Singh, Brian Kirkpatrick, Bhavin Padhiar, Aram Shemmassian, Ramnik Singh, Steve Tovar, Sophia Wu, James Jen, Katrin Kahen, Courtney Gonzalez, Grant Alkin, Bassel Rifai, Poonam Desai, Damien Wood, Robert Kwak, Mariam Guirguis, Jayantha Thiyanaratnam, Preethi Poulose, Yuli Chang, Maggie Wells, Jamie L. Marshall, Aya Pusic, Marina Stavchanskiy, Thomas Kho, Nick Garrone, Jeyling Chou, Stephanie Calvillo, Charles Hu, Emily Vollmer, Winston Wu, Henry Lin, Sumit Datta, Leo Thai, Sheila Lezcano, Devon M Huff, Santino Laxamana, Meghann Ribbens, Mara Pavel-Dinu, Yein Kim, Yibing Chen, Eunha Kim, Nneka Orjiakor, Sarah Kim, Allison Milchanowski, Edwin Paz, Manyak Saakyan, Erin Marsh, Linda Kao, Russell Powell, Christopher N.H. Bui, Nikki Villarasa, Joy Wu, Eric Paul, Artemis Deravanesian, Aria Hong, Kha Nguyen, Akanksha Chhabra, Leena Tekchandani, Toni Lee, Utpal Banerjee, William R. Marrs, Elena Bibikova, Eric Lee, Lindsay Hovestreydt, Anna L. Abrams, Jordan Fabrikant, Joseph Cao, Chad Goffstein, So-Youn Lee, Daniel Chang, Will Yarosh, Peter Lin, Kosha Soneji, Christopher Lee, Thanh Luu, Katherine Fu, Maurine Tong, Amanda O'Donahue, Amanda Abramson, Desiree Rosselli, Gemma Lewis, Farid Jamshidian, George Yen, Jiong Chen, Amir Yavari, Yasaman Moshfegh, Amit Misra, Lephong Pham, Ernesto Nodado, Tanya Minasian, Ryan Hassid, Erika Stageberg, Stacy Chan, Cynthia Nguyen, Yuki Yamauchi, Marc Uemura, Janet Lee, Helena Minye, Aileen Chang, Alex Pavlenko, Kelly Trangsrud, Phyllis Gayda, Miles Morimoto, Kirsten Regalia, Jenny Chan, Ardy Davarifar, Shahbaz Farnad, Peter Dang, Daniel Solomin, Laura Yee, Jenny Anderson, Albert Cespedes, To Hang S Lee, Julie Luu, Jessica Murray, Allen Bookatz, Keenan Bashour, Joo Choi, Adam D. Langenbacher, Emil Kohan, Amy M. Chan, Sarah Min, Melissa M. Kelley, Miriam Segura, Elsa Beyer, Sarah Phaklides, Natalie Buu, Emily Weiss, Mary Unkovic, Julie Hong, Emily Skinner, Farnaz Shadpour, Wah-Yung Tsang, Eddie Garcia, Divi Ramola, Latiffe Amado, Srpouhi Gasparyan, Sarah Brewer, Kevin Yackle, and Ndidi Onugha

Subjects
Discovery science, Genetics, General Immunology and Microbiology, QH301-705.5, General Neuroscience, Genomics, In vivo analysis, Biology, Development, Genetics/Genomics/Gene Therapy, Data science, Science education, Genome, General Biochemistry, Genetics and Molecular Biology, Community Page, ComputingMilieux_COMPUTERSANDEDUCATION, Identification (biology), Drosophila, Biology (General), General Agricultural and Biological Sciences, Curriculum, Functional genomics
Abstract

The excitement of scientific research and discovery cannot be fully conveyed by didactic lectures alone. Several recent initiatives and proposals, therefore, have supported a more participatory, discovery-based instruction for undergraduate science education [1,2]. In functional genomics, we have found an ideal platform to simultaneously benefit students and contribute to scientific discovery. The sequencing of eukaryotic genomes has facilitated the identification of complete sets of genes in humans and model genetic organisms. This has allowed many forms of high-throughput analyses of transcriptional profiles, protein interactions, structural motifs, and even genome-wide knock-downs in cell lines or in selected organisms. However, one of the best tools to provide functional information about gene action— obtaining in vivo evidence about the phenotype resulting from heritable loss of function—is difficult and less amenable to high-throughput research. We were able to achieve a large-scale in vivo analysis with a significant number of undergraduate students at UCLA, called the UCLA Undergraduate Consortium for Functional Genomics. This work, a practical manifestation of policy positions proposing discoverybased education, is described in summary form here (and in Box 1) and in detail online at http://www.bruinfly.ucla.edu. This effort combines professional-quality research with a strategy for research-based undergraduate education.

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