Resume

• 2003

  Doctor of Philosophy (Ph.D.)

  Microbiology and Immunology

  Clemson Graduate Microbiological Association\nAEL Honor Society\nPKP Honor Society

  Clemson University

• 1997

  Bachelor of Science (BS)

  Biochemistry

  Womens' Intramural Soccer

  Biology Club

  Fellowship of Christian Athletes

  College of Charleston

  Bachelor of Arts (BA)

  Chemistry

  Co-Founder & Treasurer: Clemson Graduate Microbiological Association; Alpha Epsilon Delta Graduate Honor Society

  Race for the Cure

  flag football

  College of Charleston

  Teaching Techniques Certification

• Mammalian Cell Culture

  Gel Electrophoresis

  Molecular Biology

  Animal Models

  Technical Writing

  Primary Cell Isolation

  qRT-PCR

  Confocal Microscopy

  Career Development

  Teaching

  Active Learning

  PCR Primer Design

  Research

  Dendritic Cells

  Fluorescence Microscopy

  T cells

  Flow Cytometry

  Customer Service

  Cell Culture

  Mouse Handling

  Defective chemokine signal integration in leukocytes lacking Activator of G protein Signaling 3 (AGS3)

  Joe B. Blumer

  Stephen M. Lanier

  Hyeseon Cho

  Xian-Kui Zhang

  William Gene Robichaux III

  AGS3 (Gpsm1)

  an accessory protein for G protein signaling

  has functional roles in the kidney and CNS. Here we show that AGS3 is expressed in spleen

  thymus and bone marrow-derived dendritic cells (BMDCs)

  and is upregulated upon leukocyte activation. We explored the role of AGS3 in immune cell function by characterizing chemokine receptor signaling in leukocytes from mice lacking AGS3. No obvious differences in lymphocyte subsets were observed. Interestingly

  however

  AGS3-null B- and T-lymphocytes and BMDCs exhibited significant chemotactic defects as well as reductions in chemokine-stimulated calcium mobilization and altered Erk and Akt activation. These studies indicate a role for AGS3 in the regulation of G-protein signaling in the immune system

  providing unexpected venues for the potential development of therapeutic agents that modulate immune function by targeting these regulatory mechanisms.

  Defective chemokine signal integration in leukocytes lacking Activator of G protein Signaling 3 (AGS3)

  Wei Y

  Wagner TE

  Yu X

  Li J

  Kotturi HS

  Cancer Gene Therapy

  We previously demonstrated that a novel fusion protein MULT1E/FasTI expressed by TC-1 tumor cells inhibited tumor growth by simultaneously activating NKG2D expressing cells

  such as NK cells

  through the MULT1E portion and sending a death signal into cells through the Fas portion (Kotturi et al.

  Gene Therapy

  2008). In this study

  an adenoviral gene delivery system was used to deliver this fusion protein. Our data indicate that adenoviral vector can efficiently deliver the MULT1E/FasTI fusion protein into TC-1 cells both in vitro and in vivo as assayed by RT-PCR

  FACS analysis

  caspase-3 activity and decreased in vivo tumor growth. This study further confirms that MULTE/FasTI represents a powerful bi-functional

  therapeutic protein for the treatment of cancers.

  In vitro and in vivo delivery of novel anticancer fusion protein MULT1E/FasTI via adenoviral vectors

  Tew KD

  Townsend DM

  Manevich Y

  Drake RR

  Blumer JB

  Jones EE

  Reyes L

  Gao P

  Ye ZW

  Zhang J

  PLoS One

  To interrogate why redox homeostasis and glutathione S-transferase P (GSTP) are important in regulating bone marrow cell proliferation and migration

  we isolated crude bone marrow

  lineage negative and bone marrow derived-dendritic cells (BMDDCs) from both wild type (WT) and knockout (Gstp1/p2(-/-)) mice. Comparison of the two strains showed distinct thiol expression patterns. WT had higher baseline and reactive oxygen species-induced levels of S-glutathionylated proteins

  some of which (sarco-endoplasmic reticulum Ca2(+)-ATPase) regulate Ca(2+) fluxes and subsequently influence proliferation and migration. Redox status is also a crucial determinant in the regulation of the chemokine system. CXCL12 chemotactic response was stronger in WT cells

  with commensurate alterations in plasma membrane polarization/permeability and intracellular calcium fluxes; activities of the downstream kinases

  ERK and Akt were also higher in WT. In addition

  expression levels of the chemokine receptor CXCR4 and its associated phosphatase

  SHP-2

  were higher in WT. Inhibition of CXCR4 or SHP2 decreased the extent of CXCL12-induced migration in WT BMDDCs. The differential surface densities of CXCR4

  SHP-2 and inositol trisphosphate receptor in WT and Gstp1/p2(-/-) cells correlated with the differential CXCR4 functional activities

  as measured by the extent of chemokine-induced directional migration and differences in intracellular signaling. These observed differences contribute to our understanding of how genetic ablation of GSTP causes different levels of myeloproliferation and migration

  Glutathione S-transferase P influences redox and migration pathways in bone marrow

  Wei Y

  Wagner TE

  Yu X

  Li J

  Kotturi HS

  Gene Therapy

  Tumor cells evade immunosurveillance by elements of the innate immune system

  such as natural killer (NK) cells

  by downregulating or 'shedding' certain cell-surface molecules like mouse UL16-binding protein-like transcript 1 (MULT1) that can activate NK cells through NK cell receptors such as NKG2D; they also avoid Fas-mediated apoptosis by downregulating its expression. In the present study we report the design and evaluation of the antitumor activity of a novel fusion protein

  MULT1E/FasTI

  consisting of the extracellular domain of MULT1 and the transmembrane and intracellular domains of Fas. The fusion construct (pMULT1E/FasTI) was transfected into the mouse pulmonary carcinoma cell line TC-1; and stable cell clones expressing the fusion protein were established. In-vitro cell culture studies demonstrated that the binding of the NKG2D/Fc

  a recombinant protein of mouse NK cell receptor

  to MULT1E/FasTI expressed on tumor cells was able to elicit apoptosis as assayed by Annexin V-fluorescein isothiocyanate staining and caspase-3 enzyme-linked immunosorbent assay and to activate NKG2D-expressing cells

  such as NK cells. In-vivo subcutaneous tumor studies demonstrated that tumor cells expressing MULT1E/FasTI grew significantly slower than cells without the protein. Pulmonary metastasis studies showed that most of the mice completely rejected tumor cells expressing MULT1E/FasTI. This approach may generate a new therapeutic agent for tumor treatment when combined with tumor cell-specific gene delivery vehicles such as oncolytic adenovirus vectors.

  Tumor cells expressing a fusion protein of MULT1 and Fas are rejected in vivo by apoptosis and NK cell activation

  Eblen ST

  Lopez-Berestein G

  Rodriguez-Aguayo C

  Kistner-Griffin E

  Liu Y

  Blumer JB

  Sakati W

  Zheng H Ziebarth A

  Amber A. Bradley

  Carcinogenesis

  The E3 ubiquitin ligase EDD is overexpressed in recurrent

  platinum-resistant ovarian cancers

  suggesting a role in tumor survival and/or platinum resistance. EDD knockdown by small interfering RNA (siRNA) induced apoptosis in A2780ip2

  OVCAR5 and ES-2 ovarian cancer cells

  correlating with loss of the prosurvival protein myeloid cell leukemia sequence 1 (Mcl-1) through a glycogen synthase kinase 3 beta-independent mechanism. SiRNA to EDD or Mcl-1 induced comparable levels of apoptosis in A2780ip2 and ES-2 cells. Stable overexpression of Mcl-1 protected cells from apoptosis following EDD knockdown

  accompanied by a loss of endogenous

  but not exogenous

  Mcl-1 protein

  suggesting that EDD regulated Mcl-1 synthesis. Indeed

  EDD knockdown induced a 1.87-fold decrease in Mcl-1 messenger RNA and EDD transfection enhanced murine Mcl-1 promoter-driven luciferase expression 5-fold. To separate EDD survival and potential cisplatin resistance functions

  we generated EDD shRNA stable cell lines that could survive initial EDD knockdown and showed that these cells were 4- to 21-fold more sensitive to cisplatin. Moreover

  transient EDD overexpression in COS-7 cells was sufficient to promote cisplatin resistance 2.4-fold

  dependent upon its E3 ligase activity. In vivo

  mouse intraperitoneal ES-2 and A2780ip2 xenograft experiments showed that mice treated with EDD siRNA by nanoliposomal delivery [1

  2-dioleoyl-sn-glycero-3-phophatidylcholine (DOPC)] and cisplatin had significantly less tumor burden than those treated with control siRNA/DOPC alone (ES-2

  77.9% reduction

  P = 0.004; A2780ip2

  75.9% reduction

  P = 0.042) or control siRNA/DOPC with cisplatin in ES-2 (64.4% reduction

  P = 0.035)

  with a trend in A2780ip2 (60.3% reduction

  P = 0.168). These results identify EDD as a dual regulator of cell survival and cisplatin resistance and suggest that EDD is a therapeutic target for ovarian cancer.

  EDD enhances cell survival and cisplatin resistance and is a therapeutic target for epithelial ovarian cancer

  Y Wei

  TE Wagner

  X Yu

  HS Kotturi

  J Li

  Dendritic cell-mediated cancer immunotherapy employs several ways to engage tumor antigens. We have demonstrated in both pre-clinical animal studies and early clinical trials that dendritomas

  highly purified hybrids between dendritic cells (DC) and tumor cells

  are superior activators of anti-tumor immunity. It has been argued

  however

  that DC vaccines may be dysfunctional in lymph node migration. In the present study we examined inflammatory chemokine and chemokine receptor expression as well as other maturation induced genes in dendritomas produced from either immature or mature DCs in order to shed light on their capacity to migrate from injection sites to draining lymph nodes and elicit an appropriate immune response. RNA microarray analysis was used to identify gene expression profiles for inflammatory chemokines and receptors and other maturation induced genes within dendritomas

  lysate-pulsed dendritic cells

  immature DCs and mature DCs. Gene regulation was confirmed with relative quantification

  real-time RT-PCR in a separate experiment. We found that fusion of immature DCs to tumor cells initiates maturation with respect to inflammatory chemokines

  chemokine receptors and other maturation induced genes in a similar pattern as LPS matured DCs. Interestingly

  we saw a reversed gene profile when mature DCs were fused to tumor cells. LPS matured DCs displayed the chemokine repertoire expected with DC maturation; however

  once fused to tumor cells

  these chemokines and other maturation induced genes reverted to levels comparable to immature DCs. It appears that mature DCs used for dendritoma production result in a de-mature genotype. Our results indicate that dendritomas from immature DC/tumor cell fusions may be more effective in migration from injection site to draining lymph nodes and

  therefore

  would be more effective in stimulating anti-tumor immunity.

  Fusion induced reversal of dendritic cell maturation: an altered expression of inflammatory chemokine and chemokine receptors in dendritomas

  Examining the interactions of AGS3

  Gai2 and chemokine receptors CXCR4 and CCR7 after stimulation with agonist. Our lab has developed a mouse model lacking the AGS3 protein and I am investigating the functional defects in chemokine signaling

  calcium mobilization

  Erk and Akt activation

  and protein recruitment in mice lacking this protein compared to wild type mice.\n\nDelineating signaling events downstream of GPCRs leads to discovery of novel drug targets. The basic science behind understanding these interactions is critical as greater than 30% of all pharmaceutical drugs target or involved GPCRs.

  O'Connor

  Branham

  College of Charleston

  University of Maryland University College

  Greenville Hospital

  Oncology Research Institute

  Pier 1 Imports

  The Citadel

  Charleston Southern University

  Medical University of South Carolina

  Clemson University

  ASPET

  The Home Depot

  The ASPET Washington Fellow program enables early career scientists interested in science policy the opportunity to learn about and become more engaged in public policy issues. Fellows engage in advocacy efforts both on Capitol Hill and in their home districts.

  ASPET

  Pier 1 Imports

  Assistant Manager

  Charleston

  South Carolina Area

  Collegiate Traveling Faculty in Biological Sciences\nCourses: Human Anatomy and Physiology I & II

  Microbiology

  Introduction to Biology

  Human Biology

  Nutrition with corresponding labs\n\nOne of two CTF representatives on the world-wide UMUC Faculty Advisory Committee

  Assistant Professor

  US Military Posts in Europe

  University of Maryland University College

  Clemson

  SC

  Teaching Assistant

  Clemson University

  Charleston

  SC

  Seasonal Success Captain & Sales Associate

  The Home Depot

  PACD Fellow

  mentored teaching experience at a primarily undergraduate institution. NIGMS SC INBRE Grant for Postdoctoral Academic Career Development (PACD).

  The Citadel

  Graduate Research Assistant

  Culture of primary dendritic cells from murine bone marrow

  production of dendritomas (fusion of dendritic cells with tumor cells)

  PEG fusion

  electroporation

  tritium thymidine incorporation cell proliferation assays

  irradiation of dendritomas

  certified GMP in accordance with working in a clinical trial laboratory.

  Greenville Hospital

  Oncology Research Institute

  Post Doctoral Fellow

  Self-directed design and implementation of primary research project: The Characterization of AGS3 and AGS4 in Primary Immune Cells Utilizing Both AGS3- and AGS4-null Mouse Models.\n\nResearch techniques routinely performed include

  primary murine cell culture

  culture and maintenance of cell lines (both murine and human)

  primary cell isolation & culture including dendritic cells

  B cells

  T cells

  and neutrophils

  immune assays

  transwell chemotaxis assays

  calcium mobilization assays

  western blotting/immunoblotting

  blotting for phosphorylated proteins

  genotyping

  real-time PCR

  signal transduction assays

  bioluminescent resonance energy transfer (BRET)

  FRET

  light-field microscopy

  fluorescence microscopy

  confocal microscopy

  immunocytochemistry (ICC)

  immunohistochemistry (IHC)\n\nLab management responsibilities including Marketplace ordering

  maintenance of mouse colonies

  training of graduate student rotations and lab technicians

  and maintenance of routine OSHA/Biohazard safety operations consistent with a Biosafety Level-2 laboratory. Work with E. coli

  pertussis toxin

  endotoxin

  and other BSL-1/2 toxins and/or biohazards were routinely performed in accordance with federal regulations.

  Medical University of South Carolina

  Adjunct Instructor

  Instructor of Cell and Molecular Biology for science majors.

  College of Charleston

  Charleston Southern University

  Charleston

  South Carolina Area

  Assistant Professor

  Washington Fellow

  ASPET

  GSPAC member

  ASBMB

  Chair

  Career Development Committee

  MUSC PDA