Program Overview:   10 week program MD Anderson Cancer Center in Houston, Texas Students will be matched with a faculty mentor in any number of areas of biomedical research where a project will be assigned. Participants work alongside the mentor in a lab or clinic on projects designed by faculty to reflect current research. Workshops and lectures provide opportunities to connect with faculty, residents, postdoctoral and clinical fellows, and other participants. Through the program, students assess goals related to careers in oncology research and patient care. Symposium in which participants present talks and posters on their research projects to peers and faculty

Fortoul, Marla

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BIOLOGY SUMMER INTERNSHIPMARLA FORTOUL SUMMER UNDERGRADUATE PROGRAM AT MD ANDERSON CANCER CENTER 10 weeks program  MD Anderson Cancer Center in Houston, Texas.  Students will be matched with a faculty mentor in any number of areas of biomedical research where a project will be assigned. Participants work alongside the mentor in a lab or clinic on projects designed by faculty to reflect current research.  Workshops and lectures provide opportunities to connect with faculty, residents, postdoctoral and clinical fellows, and other participants.  Through the program, students assess goals related to careers in oncology research and patient care.  Symposium in which participants present talks and posters on their research projects to peers and faculty. WHY SHOULD YOU GO TO MD ANDERSON ? Discover Possible Career Paths Networking opportunities Learning multiple techniques Shadowing Physicians  Oncology  PhD MD/PhD  Workshops Simulation Center  Speech Elevator  Cancer  Meet Students from all over the country  Learn about advancements in science Experience for Resume Research and clinical  Being at MD Anderson  Best Hospital in Cancer Care Groundbreaking research  Innovative Clinical Care Clinical Trials  Patients travel all over the world to be treated at MD AndersonPROJECTDETERMINE THE EFFECT OF IBRUTINIB ON HUMAN T CELLS. Department of Stem Cell Transplantation PI: Qing Ma, PhD Dan Li, Sr. ResearcherIBRUTINIB Therapy for B-cell malignancies Chronic Lymphocytic Leukemia:  It develops slowly, creating defective white blood cells that can’t produce antibodies against infection.  The average age of patients at diagnosis is 72.  Most Common type of Leukemia  Effects of Ibrutinib:  Inhibits Bruton’sTyrosine Kinase (BTK) Blocks downstream B cell receptor activation. Investigating the drug’s effect on Inducible T cell Kinase (ITK) Determine the effect on T cell receptor and T cell proliferation. PROJECTS Human T-Cell Activation  Human CD4 T Cell Proliferation  Human T Cell Counts Inhibited in the Presence of Ibrutinib Human CD4 T cell Proliferation in the Presence of IbrutinibPOSTERIntroductionIbrutinib, is an orally active agent, whichworks by covalent irreversible binding ofCys-481 in the kinase domain of Bruton’sTyrosine Kinase (BTK).1-2 BTK is a non-receptor tyrosine kinase of the Tec-kinasefamily and is of central importance in B-cellreceptor (BCR) signaling. 3In addition to BTK inhibition, it has beenhypothesized Ibrutinib as a target ofInterleukin-2 Inducible Kinase (ITK) due tosignificant homology with BTK. ITK is a Tcell dominant member of the TEC-kinasefamily that drives proximal T-cell receptor(TCR) signaling.4 ITK is active in T-cellmalignancies and has been linked to tumorimmune invasion and survival through itscritical role in T helper type 2 (Th2) celldifferentiation, making it an attractivetherapeutic target.5ConclusionsHuman T cells can be activated invitro by using CD3+CD28 mAb. CFSEassay measured human CD4 T cellproliferation showing increased divisionwhen stimulated with CD3+CD28 mAb.Furthermore, subsets of human T-cells,CD4 and CD8, can be identified usingsurface markers. Also, subsets of CD4 andCD8 are positive for CD45RA and CCR7staining markers. These markers allow theidentification of naïve, central memory,effector memory, and TEMRA CD4 andCD8 human T cells.It was identified a reduced number ofactivated human T cells in vitro as Ibrutinibconcentration increased until it reached 5µM which lead to toxicity for the cellsresulting in reduced number of cells andincreased cell death. CFSE assaymeasured human CD4 T cells proliferationin the presence of Ibrutinib, it showed aninhibition of CD4 T cells. The proliferationindex of CD4 T cells decreased as theconcentration of Ibrutinib increased, itdecreased exponentially after 1 µM.In conclusion, Ibrutinib inhibits humanT cells proliferation, in particular CD4 Tcells. The inhibition is dose dependent andstarts becoming toxic for the cells at 5 µM.Title of the Poster Presentation Goes HereMarla Fortoul1; Dan, Li2; Qing, Ma, PhD3;1[MD Anderson Summer Undergraduate Program], 2&3 [MD Anderson Cancer Center Department of Stem Cell Transplantation-Research]Ibrutinib Inhibits Huma  CD4 T Cell P oliferation l  Fortoul1; Dan Li2; Qing Ma, PhD2;1[  nderson Summer Undergraduate Program], 2 [MD Anderson Cancer Center Department of Stem Cell Transplantation-Research]References1. Herman, S. E., C. U. Niemann, M. Farooqui, J.Jones, R. Z. Mustafa, A. Lipsky, N. Saba, S.Martyr, et al. 2014. Ibrutinib-inducedlymphocytosis in patients with chronic lymphocyticleukemia: correlative analyses from a phase IIstudy. Leukemia 28: 2188–2196.2. Schwarzbich MA., Witzens-Harig M. (2014)Ibrutinib. In: Martens U. (eds) Small Molecules inOncology. Recent Results in Cancer Research,vol 201. Springer, Berlin, Heidelberg3. Burger, J. A., and J. J. Buggy. 2013. Brutontyrosine kinase inhibitor ibrutinib (PCI-32765).Leuk. Lymphoma 54: 2385–2391.4. Berg LJ, Finkelstein LD, Lucas JA, SchwartzbergPL. Tec family kinases in T lymphocytedevelopment and function. Annu Rev Immunol.2005;23:549-600.5. Dubovsky, J.A., Beckwith, K.A., Natarajan, G.,Woyach, J.A., Jaglowski, S., Zhong, Y., Hessler,J.D., Liu, T.M., Larkin, K.M., et al. 2013. Ibrutinib isan irreversible molecular inhibitor of ITK driving aTh1-selective pressure in T lymphocytes. Blood122:2539-2549.Materials and Methods• PBMCs IsolationPeripheral blood mononuclear cells(PBMCs) were isolated via density gradientcentrifugation over Ficoll-Paque from buffy coatpreparations derived from whole blood of healthyvolunteer donors (Gulf Coast Regional BloodCenter, Houston, TX).• Tissue CulturePBMCs were cultured in a 96-well V-bottom plate in culture media (RPMI 1640, 10%FBS, and 1% P/S) at 0.2 x 106 cells/well. Culturedcells were stimulated using 1 µg/mL anti-CD3 and 1µg/mL anti-CD28 mAb for 6 days.• CFSE ProliferationA CellTrace CFSE Stock thawed anddiluted in phosphate-buffered saline (PBS). T cellswere resuspended in 1 mL of PBS with CFSE andincubated for 2 minutes at room temperature. Cellswere washed and resuspended in culture media tostabilize CFSE staining to track cell division.• IbrutinibAn Ibrutinib stock concentration of 100mM was diluted into different concentrations (0, 0.5,1, 5, and 10 µM) using DMSO. 1 µL of differentconcentrations of Ibrutinib were added to PBMCscell culture.• Flow CytometryCultured T cells were stained withmonoclonal antibodies (CD4-PE, CD8-APC, CFSE-FITC, CCR7-Pe-Cy, and CD45RA-PB) Sampleswere acquired on Canto II Analyzer usingFluorescence-activated cell sorting and analyzedwith FlowJo.Human T-Cell ActivationFigure 1. Human T Cell Activation.Human PBMCs were stimulated withCD3+CD28 Ab for 6 days. They werestained for expression of CD4, CD8,CCR7, and CD45RA. Results depict (a)SSC and FSC plot with gated livelymphocytes, (b) contour plot ofexpression of gated CD4 and CD8 fromlive lymphocytes population, (c) CD4 andCD8 gated cells were separated into foursubsets (CM= central memory, N=Naive,TEMRA= terminal differentiated, and EM=effector memory) based on CD45RA andCCR7 labeling. (d) Column graphscomparing Subsets of CD4 and CD8 Tcells between stimulated andunstimulated T cells.BHuman CD4 T Cell Proliferation Figure 2. Human T Cell Proliferationusing CFSE assay. CFSE assay CD4 Tcells stimulated with CD3+CD28 mABfor 6 days. (a) Unstimulated CD4 cellswith 1 generation. (b) Stimulated CD4cells with 6 generations.Results Human T Cell Counts Inhibited in the Presence of IbrutinibHuman CD4 T cell Proliferation in the Presence of IbrutinibObjectivesDetermine the effect of Ibrutinib on human Tcells.Figure 3. Human T Cell Inhibited in thePresence of Ibrutinib. CFSE labeled Tcells stimulated with CD3+CD28 Ab for 6days with different concentrations ofIbrutinib (0, 0.5, 1, 5, and 10 µM). (a)Results depict SSC and FSC plot withgated live lymphocytes for unstimulated andstimulated cells with various concentrationsof Ibrutinib. (b) Column graph showingnumber of lymphocytes in each group.Contact InformationMarla FortoulNova Southeastern University, SURP at MD Andersonmarlacarolinafortoul1997@gmail.comFigure 4. Human CD4 T cellProliferation in the Presence ofIbrutinib. CFSE labeled T-cells stimulatedwith CD3+CD28 for 6 days with differentconcentrations of Ibrutinib (0, 0.5, 1, 5, and10 µM). Results depict a histogramshowing the proliferation of CD4 cells. (B)Column Graph depicting the division indexfor unstimulated and stimulated CD4 cellswith different concentrations of Ibrutinib.5 µM BA020406080100CM Naïve TEMRA EMUS S020406080CM Naïve TEMRA EMUS SDStimulatedACBFSCSSCCD8CD4UnstimulatedCD45RACCR7CD4ACBCD45RACCR7CD8FSCSSCCD8CD4CD4 CD8CD45RACCR7CD45RACCR7CD4 CD8NCMEMEMCM NAUnstimulatedCFSE# CellsStimulatedCFSE# CellsCM CM NNEMEMTEMRA TEMRATEMRATEMRA0 µM10 µM1 µM0.5 µM5 µMFSCSSCFSCSSCFSCSSCFSCSSCFSCSSCBUnstimulatedFSCSSCUnstimulated 0 µM1 µM0.5 µMCFSE# CellsCFSE# CellsCFSE# CellsCFSE# CellsCFSE# CellsAPPLICATION REQUIREMENTS Nova Southeastern registered Halmos College junior or senior for fall 2019 with: Science GPA 3.0+ and cumulative GPA 3.0+. Successfully completed: Anatomy & Physiology, Medical Terminology, and Statistics. Preference to students with upper-level courses. Complete application submitted by October 26, 2018 including: Transcript or the CAPP report equivalent Personal Statement 3 Letters of Recommendation Application can be found online at https://cnso.nova.edu/biology/index.html under Summer Internships. If qualified, committee interview. Contact Information:• Dr. Mark Jaffe, mjaffe@nova.edu• Dr. Deanne Roopnarine, roopnari@nova.edu• Dr. Aarti Raja, araja@nova.edu.• Marla Fortoul, mf1612@mynsu.nova.edu

The University of Texas MD Anderson Cancer Center - Biology Summer Internship

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The University of Texas MD Anderson Cancer Center - Biology Summer Internship

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