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Construction and testing of chimeric antigen receptor targeting CS1 for treatment of primary effusion lymphoma
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Construction and testing of chimeric antigen receptor targeting CS1 for treatment of primary effusion lymphoma
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Content
Construction and Testing of Chimeric Antigen Receptor targeting CSl for Treatment of Primary
effusion Lymphoma
By
Songjie Gong
A Thesis Presented to the Faculty of the Graduate School
UNIVERSITY OF SOUTHERN CALIFORNIA
MASTER OF SCIENCE
IN
(MOLECULAR MICROBIOLOGY & IMMUNOLOGY)
August2015
1
II. INDEX
I. Title ............................................................................................................................... Page 1
ll.~x ............................................................................................................................ ~2
Ill. Introduction .......................................................................................................................... Page 3-8
Primary effusion lymphoma .................................................................................................... 3-4
Multiple myeloma .................................................................................................... 4-5
Cell surface glycoprotein CS 1 ...................................................................................................... 6
Anti-CS 1 monoclonal antibody ...................................................................................................... 6
Chimeric antigen receptors .................................................................................................... 6-8
IV. Materials and Methods ...................................................................................................... Page 9-11
Cells ........................................................................................................................ 9
Generation ofCAR-Anti-CSl lentiviral construct ................................................................... 9
Virus production
Gene transduction
Detection of CAR
...................................................................................................................... 10
................................................................................................................ 10-11
...................................................................................................................... 11
V. Results .................................................................................................................... Page 12-16
Generation ofCAR-Anti-CSl lentiviral construct ............................................................... 12
Efficient generation ofNK92-MI cells expressingAnti-CSl-CAR .................................... 13-15
In vitro cytotoxicity assay .................................................................................................... 15-16
VI. Discussion ..................................................................................................................... Page 17-18
VII. References ..................................................................................................................... Page 19-20
2
Ill INTRODUCTION
Primary effusion lymphoma
Primary effusion lymphoma (PEL) is a rare B-cell non-Hodgkin lymphoma presenting with a malignant
effusion without a tumor mass [3]. Patients commonly present with dyspnea (from pleural or pericardia!
disease) or abdominal distension (from peritoneal disease) [3]. PEL is always associated with HIN-8,
also known as Kaposi sarcoma-associated herpesvirus (KSHV), and is most frequently found in
immunodeficient patients, especially those with advanced AIDS [1][2].
Table 1. Comparison of characteristics of differe nt histological types of AlDS-related non-Hodgkin 's lymphomas (ARLs)
Typical CD4 count
Histological type % ofARL a t diagnosis EBV+
Burkitt's or Burkitt's-like 25%- 30% Usually normal 30%
Diffuse large B-cell (centroblastic) 25%-30% < 100/ul 40%
Diffuse large B-cell (immunoblastic) including PCNSL 25%-30% < 100/ul 90%- 100%a
Plasrnablastic lymphoma < 5% Decreased > 50%
b
Primary effusion lymphoma < 5% Decreased 90% 100%
Plasmablastic lymphoma includes HHV -8- positive disease arising from multicentric Castleman's disease and HHV-8-
negative plasmablastic lymphoma of the oral cavity.
•Primary centra l nervous system lymphoma in the HIV setting has a 100% EBY infection rate.
bPlasmablastic lymphoma of the oral cavity is typically HHV-8 negative by immunohistochemical staining for latency associated nuc lear antigen- I, but HHV-8 DNA is detectable by polymerase chain reaction in many cases (68).
Abbreviations: EBY, Epstein- Barr virus; HHV-8, human herpesvirus-8; PCNSL, primary central nervous system
lymphoma.
Fig.1 Comparison of characteristics of different histological types of AIDS-related non-Hodgkin's lymphomas
(ARLs) (Ephraim Hochberg, 2007)
PEL is usually extremely aggressive and the lymphoma cells can be propagated to fairly distant sites.
HIV-related complications and opportunistic infections are often fatal. In the rare HIV-negative cases,
the outcome may be better. To diagnose PEL, an infection with HHV -8 must be present. HIN-8 in tissue
samples can be detected by a latency-associated nuclear antigen- I (LANA-1) assay. PEL is usually
resisted to conventional chemotherapy. Currently the therapeutic options for PEL usually combine
3
CHOP ( cyclophosphamide, doxorubicin, vincristine, and prednisone ), chemotherapy and antiretroviral
therapy (in case of HIV-positive individuals) [3]. Even with these complicated treatments regimens,
patients have a median survival time of only approximately 6 months. As such, there is an urgent need
for novel targeted therapies for PEL
Multiple myeloma
Multiple myeloma is a cancer that begins in plasma cells, a type of white blood cell derived from B
lymphocytes [4]. These cells are part of our immune system, protecting our body from being attacked by
germs, viruses and other harmful substances. B lymphocytes are produced in the bone marrow, and then
move to the lymph nodes. Multiple myeloma develops in B lymphocytes after they have left the
germinal center. Myeloma cells accumulate in bones and bone marrow. They may cause damage to solid
parts of bones, leading to weakness and fracture of the bones.
4
Red marrow
where plasma
cells are made
Multiple Myeloma
Normal plasma cells
t ~:::( ~A t'b d'
::f."
1 1
~./. n 1 o 1es
' ¥ 1 r
\1
Multiple myeloma cells (abnormal plasma cells)
© 2014 Terese Winslow LLC
U.S. Govt has certain rights
Fig.2 Multiple myeloma cells are abnormal plasma cells (a type of white blood cell) that build up in the
bone marrow and form tumors in many bones of the body. Normal plasma cells make antibodies to help
the body fight infection and disease. As the number of multiple myeloma cells increases, more
antibodies are made. This can cause the blood to thicken and keep the bone marrow from making
enough healthy blood cells. Multiple myeloma cells also damage and weaken the bone. (National Cancer
Institute, General Information about Plasma Cell Neoplasms, 2015)
Currently treatment for multiple myeloma is focused on therapies to decrease the clonal plasma cell
population and thus decrease the signs and symptoms of disease [7]. Although several drug therapies are
available for multiple myeloma, it still remains incurable.
5
Cell surface glycoprotein CSl
In order to develop new immunotherapies for the treatment of multiple myeloma, cell surface
glycoprotein CSI (CD2 subset-I, CRACC, SLAMF7, and CD319), a member of the signaling
lymphocyte activating-molecule-related receptor family has been identified [8]. CSI is highly
selectively and consistently expressed on myeloma cells but is generally not expressed by normal tissues
or stem cells [9][12]. Additionally, the amplifications of chromosome lq, where CSI gene is located,
can be found frequently in aggressive myeloma and has been linked to early myeloma-related death
due to over expression of the cell cycle regulator CKSIB [IO].
Anti-CSl monoclonal antibody
A humanized monoclonal antibody (mAb) that specifically targets CS 1 has been demonstrated to
possess potent in vitro and in vivo activity against myeloma cells [ll ]. Via NK-mediated
antibody-dependent cellular cytotoxicity (ADCC), this antibody can partly, even mostly, eliminate
myeloma cells, providing the preclinical framework of targeting CSl to improve the cure rate of patients
in multiple myeloma [ll]. Thus, targeting CSl with Anti-CS! mAb has potential for the treatment of
multiple myeloma.
Chimeric antigen receptors
Chimeric antigen receptors (CARs ), also known as artificial T cell receptors, allow modification of
extracted patient T-cells that improve on their existing recognition abilities (immunotherapy).
"First-generation" CARs had the intracellular domain from the CD3 t;- chain, which is the primary
6
transmitter of signals from endogenous TCRs. "Second-generation" CARs add intracellular signaling
domains (e.g. from 4-lBB and CD28) to provide additional signals to the T cell, which improves the
SW'vival and antitumor activity of T cells. 'Third-generation" CARs have combined multiple signaling
domains [13). The T cell costimulatory molecules, such as 4-lBB and CD28, c1itically regulate the
survival of activated and memory CDS T cells, while the ~ chain of CD3~ domain plays an important
role in recognition to several intracellular signal-transduction pathways which can increase the
efficiency of immune response [15).
Chimeric Antigen Receptors
_,
mAb 3rd Generation CAR-Tcell
-- CD31; ~CD28 - 4-1BB
# TCR
Fig.3 Chimeric antigen receptors. Chimeric antigen receptors (CARs) combine antigen-recognizing
variable regions (scFVs) from monoclonal antibodies (mAb) with intracellular T-cell signaling domains.
scFv region is derived from vL and vH region of an antibody and binds the target antigen.
Artificial T cell receptors are a potential therapy for cancer, which utilizes a technique called adoptive
cell transfer [14). T cells are extracted from a patient and modified so that they can express receptors
specific to the cancer. The T cells are then reintroduced into the patient for the T cells to recognize and
specifically eliminate the cancer cells. The first generation of CAR-modified T cells have shown some
7
promising results in pre-clinical trials. Second and third generation CAR-modified T cells are also
effective in amplifying activation signals, helping proliferation, cytokines production and effector
function in pre-clinical trials. Both the second and the third generation CAR-modified T cells are
entering clinical trials.
8
IV. MATERIALS AND METHODS
Cells
Primary effusion lymphoma (PEL) cells, BCBLl and JSCl, and HEK-293FT cells were available in our
laboratory. PEL cells were maintained in RPMI medium supplemented with 20% (v/v) fetal bovine
serum (FBS) and 1 % antibiotic-antimycotic solution. NK-92MI, an interleukin-2 (IL-2) independent
Natural Killer Cell line derived from the NK-92, was obtained from ATCC (American Type Culture
Collection). The base medium, Alpha Minimum Essential medium without ribonucleoside and
deoxyribonucleoside, was supplemented with 2mM L-Glutamine and l.5g/L sodium bicarbonate, 0.2mM
inositol, O. lmM 2-mercaptoethanol, 0.02 mM folic acid, horse serum to a final concentration of 12.5%,
fetal bovine serum to a final concentration of 12.5% to culture NK-92MI cells.
Generation of CAR-Anti-CSl lentiviral construct
The Anti-CS 1-scFv fragment was cloned into a CAR vector designed in our laboratory
(pLenti-EFlu-scFv-MYC-hCD8-h41BB-CD3Z-T2A-EGFP). Both the Anti-CSl-scFv fragment and
vector were digested with Nhel and Mlul restriction enzyme to replace existing scFv fragment with the
anti-CSl fragment and to generate a pLenti-EFlu-Anti-CSl-MYC-hCD8-h41BB-CD3Z-T2A-EGFP
construct. Besides this CAR construct, a pLenti-EFlu-Anti-CSl-MYC-hCD8-CD3Z-T2A-EGFP
construct, which lacked the 4 lBB costimulatory domain, was also generated.
9
Virus production
To generate Anti-CSl-CAR lentivirus, we used calcium phosphate DNA precipitation to transfect
HEK-293T cells. HEK-293T cells were purchased from ATCC. 293T cells were maintained in 10 cm
tissue culture dishes (Falcon, Becton Dickinson) for 24 hand transfected with lOµg of Anti-CSl-CAR
plasmid along with 7.5 µg of PSPAX and 2µg of PLP/VSVG packaging plasmids. After 24 hours, the
medium was replaced with fresh DMEM with 10% FBS and antibiotics. Conditioned medium
containing lentivirus was harvested 48 and 72 h after transfection, immediately filtered through a
0.45µm filter, concentrated by ultracentrifugation at 18,500 rpm at 4°C for 2 hours and re-suspended in
NK-92 cell medium to concentrate virus by 10 fold. The concentrated virus was frozen immediately in
small aliquots at -80°C until use.
Gene transduction
NK92-MI cells were plated at a density of 0.5Xl0
6
cells per well in a 6-well plate in a total volume of 2
mL, then transduced by adding 300µ1 concentrated viral supernatants and 8 µg/mL polybrene by
spinfection. For spinfection, cells were centrifuged at 2,800 rpm at 32°C for 90 min. After centrifugation,
cells were left undisturbed for 24h in a humidified incubator at 37°C, 5% C0
2
. The transduction
procedure was repeated on two successive days. Cells were then washed with NK92 culture medium and
maintained in NK-92MI culture medium to expand the cells for sorting. One week after expanding the
cells, the cells were washed 3 times with PBS and sorted based on GFP expression by flow cytometry on
a FACScan (BD Biosciences). After separating GFP-positive CAR-Anti-CSl cells, fresh NK92 culture
medium supplemented with interleukin-2 (IL-2, lµl/ml) and ciprofloxacin was added to the cells. The
10
cells were then plated in 96-well plate for initial cell culturing at 37°C. One week later, cells were
transferred to a 24-well plate with same medium conditioned with IL2 and ciprofloxacin. The cells
were expanded further in cell culture flasks without adding exogenous IL-2 or ciprofloxacin.
Detection of CAR
NK-92MI-Anti-CS1-CAR cells were stained with anti-human Myc antibody conjugated to APC
(Allophycocyanin) (Myc-APC) to detect the surface expression of Myc-tagged Anti-CSl scFV
containing CAR. These NK92-CAR cells also have GFP tag. Both, GFP and Myc-APC stains were
detected by FACSVerse flow cytometer (Becton Dickinson). Briefly, one million
NK-92MI-Anti-CS1-CAR cells or NK-92MI parental cells were washed with PBS-BSA and stained
with 5µ1 of Myc-APC antibody for 1 hour at 4°C in dark. After one hour of incubation, the cells were
washed with PBS-BSA two times and re-suspended in 0.5 ml of PBS to acquire on flow cytometer.
11
V.RESULTS
Generation of CAR- Anti-CSl lentiviral construct
To make a CAR construct, a cDNA fragment encoding CD8 signal peptide and a
single-chain-variable-fragment (scFv) fragment derived from an antibody against CSl was cloned in a
lentiviral vector in frame with a Myc epitope tag, a CD8 hinge and transmembrane domain, the
cytoplasmic signaling domain derived from human 4-lBB and signaling domain of hCD3s chain, a T2A
ribosomal skipping sequence and an enhanced green fluorescence protein (Fig. 4).
Nhe
L
scFV
~·
Digestion
with Nhe
& Miu,
EFla
:J
Ligation
Signal Peptide
(~bp)
EFla
h-'·IUB
Nhel
scFV
Mlu1 hCDS (l26 bp) hCD3~
(207 bp) (339 bp
(800bp)
BBZ
Signal Peptide
(~bp) MYC tag c391ip1
er::::
~
LI')
Digestion with Nhe & Miu,
h4-IHB
Mlu1 hCDS 1
126
bp) hCD3;
(207 bp) (339 bp/
BBZ
MYC tag t39bpJ
(63 bp)
TIA
I
Xbal::Spol
EGFPter
(717 bp)
SaA
(63bp)
T2A
SaA
EGFPter
(717 bp)
Xbal::Spol
Fig 4. A schematic
representation of the
pLenti-EFl a-Anti-CS 1-My
c-hCD8-h4-1BB-CD3Z-T2
A-EGFP construct.
The plasmid construct designated as pLenti-EFla-scFv-MYC-hCD8-h4-lBB-CD3Z-T2A-EGFP was
used as a template to replace scFv antibody region with Anti-CSl scFv fragment using Nhel and Mlul
restriction enzyme sites (Fig. 4 ). After ligation and transformation steps in cloning, screening for
12
positive clones was done by colony-PCR and DNA was prepared from positive clones by plasmid
preparation followed by confirmation of sequence by DNA sequencing (Fig. 5). The resulting construct
was designated pLenti-EFla-Anti-CS1-Myc-hCD8-h4-1BB-CD3Z-T2A-EGFP. In addition, we also
made a pLenti-EFla-Anti-CSl-l\1YC-hCD8-CD3Z-T2A-EGFP construct which lacked the h4-1BB
reg10n.
Anti-CSl CAR with h4-1BB Anti-CSl CAR without h4-1BB
Fig 5. A representative figure
showing the result of colony-PCR
screening for positive clones. Clones
shown in red frames were selected
for plasmid preparation.
Efficient generation of NK92-MI cells expressing Anti-CSl-CAR.
NK92MI cells were transduced with lentiviral supernatant from HEK-293FT cells transfected with
pLenti-EFla-anti-CS 1-l\1YC-hCD8-h4-1BB-CD3Z-T2A-EGFP vector as discussed in the Materials and
Method section. EGFP-positive cells were sorted by The F ACS Aria II cell sorter at our flow cytometry
core facility (Fig. 6).
13
NK92-MI-Anti-CS1-CAR Cells
10•
192 192
i
..
~
u
~ 128 ~ 128
Vl i
l
i
Vl
64 64
8'
i
j 10
3
:i:
~
;: 10
2
i
10
1
0 0
0 64 128 192 256 0
1
101 102 1()3 10' 105
64 128 192 256
488-FSC-Area 488-SSC-Height
488-513126-Height-Log
-
-
....
- -
-
.... . ...
, ... J17*)jJ ....
'""
mm1 10000 .11 1t
,..,
IOlUll
'""
IJJ1tll 010 ..
'"'
. ..
GFP
Fig 6. Cell sorting by flow cytometry: NK92MI-Anti-CS1-CAR cells were sorted to select
EGFP-expressing cells to expand these cells for cytotoxicity assays. Cells shown in in R3 area are
showing very high EGFP expression and were collected for further culturing and experiments.
EGFP-positive CAR expressing NK92MI cells were expanded and analyzed on a flow cytometer for
expression of CAR. Cells were gated on the live cells to exclude dead cells or cell debris. Cells stained
with anti-Myc-APC antibody were seen in upper right quadrant of dot plots as shown in Figure 7.
Double positive cells showing GFP as well as Myc-APC staining represent CAR-positive NK92MI cells.
This cytometric result clearly showed that NK92-MI cells were successfully infected with
Anti-CSl-CAR (Fig. 7).
14
1
()
(j)
(j)
All Events
,,. . .
FSC-A
·-
"":
P1
.. ..
...
mt.A
GFP-A
P1
-
...
...
1 ..
lj
..
..
~ .. ..
IK•A
APC-A
(Myc)
ii'
1
()
CL
<(
P1
GFP-A
Ul
Un-infected
Ul
Anti-CS1
Figure 7. Expression of Anti-CSl in NK92-MI cells. F1ow cytomeby clot plots illustrate expression of
Anti-CSl-Myc (Myc-APC antibody) in combination with GFP in uninfected NK92MI cells or NK92MI cells
transchiced with viral supernatant generated for CAR construct as detailed in Mateiials and Methods section.
In vitro cytotoxicity assay indicate prominent cell death of PEL cells by NK92MI-Anti-CS1-CAR
cells.
To determine whether NK92Ml-Anti-CS1-CAR cells could kill PEL cell l, NK92MI-Anti-CS1-CAR
cells expressing EGFP vector alone, full length-Anti-CSl-CAR and 4-lBB-deleted-Anti-CSl-CAR were
used as Effector (E) cells and BCBLl and JSCl PEL cell lines were used as target (T) at E:T ratio of 1:1
in a 96-well plate for 4 hours. As shown in Fig. 8, NK92MI cells expressing full length-Anti-CSl-CAR
showed prominent cell death (3.5 fold) of both the PEL cells as compared to cell death by either
NK92MI-GFP or NK92MI4-1BB-deleted-Anti-CS1-CAR effector cells suggesting that 4-lBB
15
cytosolic domain 1s required for cytotoxicity of Anti-CSl-CAR towards PEL-derived BCBL-1 and
JSC-1 cell lines.
4
Qi
~ 3.5
c
~ 3
u
:E 2.5
&.
- 2
..c
....
~ 1.5
0
= 1
QI
u 0.5
0
NK Cells+ BCBL-1 (4 hours)
4
Qi
~ 3.5
c
~ 3
u
:E 2.5
&
- 2
..c
10
Cl.I 1.5
0
1
a
0.5
0
NK Cells+ JSC-1 (4 hours)
Fig 8. NK92MI-Anti-CS1-CAR cells demonstrate antigen-specific killing of PEL cells. Following 1 month
expansion, NK92l\1I-CAR (Effector cells) were co-cultured with PEL cells (Target cells) at E:T ratio of 1 :1 for 4
hours. Results represent the fold change in target cells lysis as described in I:vlaterials and Methods section.
16
VI. DISCUSSION
Using T cells and NK cells to express a chimeric antigen receptor by genetic manipulation that is
specifically targeting some malignancies has now emerged as a promising strategy [16-18]. Ifwe would
like to produce suitable single-chain-variable-fragment to target antigen that is presented on the cell
surface of malignancies, we first need to find this specific target for scFv-CARs to bind [19]. CSl is not
expressed by normal tissues or stem cells, or only expressed under a very restricted pattern [ll].
Targeting cancer cells by CAR-CS 1 can protect normal cells from being attacked [20]. Based on what
has been discussed above, we decided to test whether CS 1 could be an ideal antigen target for treating
primary effusion lymphoma.
In this study, we successfully generated a lentiviral vector expressing CAR targeted against CS 1 using a
scFv fragment derived from a CSl monoclonal antibody. The vector contained h4-1BB costimulatory
domain and CD3t; signaling domain. Using staining with an antibody against the Myc epitope tag that
was incorporated in the extracellular domain of the CAR, we demonstrate stable expression of CAR on
the surface ofNK92-MI cells. We further show that NK92-MI cells expressing an anti-CSl-CAR vector
containing anti-CSl scFv, h4-1BB and CD3t; chain possess anti-PEL activity. In contrast, CAR lacking
the 4-lBB did not show significant killing against PEL cells, suggesting that the 4-lBB domain may
play a role in the activity of CAR. Based on these results, we plan to express anti-CSl-CAR in primary
T cells and demonstrate the activity of CS 1-CAR modified T cells to kill PEL cells in vitro and
subsequently in vivo.
17
In conclusion, we have successfully generated anti-CSl-CAR to target CSl, and have observed
remarkable cell death of PEL cells in vitro. Our study suggests that anti-CS 1 CAR may represent a
promising approach for the treatment of PEL.
18
VII. References
[1] Cesarman E, Chang Y, Moore PS, Said JW, Knowles DM (May 1995). "Kaposi's sarcoma-associated
herpesvirus-like DNA sequences in AIDS-related body-cavity-based lymphomas". N. Engl. J. Med. 332
(18): 1186-91.
[2] Staudt MR, Kanan Y, Jeong JH, Papin JF, Hines-Boykin R, Dittmer DP (July 2004). "The tumor
microenviromnent controls primary effusion lymphoma growth in vivo". Cancer Res. 64 (14): 4790-9.
[3] Chen YB, Rahemtullah A, Hochberg E. Primary effusion lymphoma. Oncologist 2007;12:569-76.
[4] Raab MS, Podar K, Breitkreutz I, Richardson PG, Anderson KC (July 2009). "Multiple myeloma".
Lancet 374 (9686): 324-39.
[5] Federico Caligaris-Cappio; Manlio Ferrarini (1997). Human B Cell Populations. Chemical
Immunology 67. Switzerland: S. Karger AG. p. 105.
[6] Tricot G (2000). "New insights into role of microenviromnent in multiple myeloma.". Lancet 355
(9200): 248-50.
[7] Korde N, Kristinsson SY, Landgren 0 (2011). "Monoclonal gammopathy of undetermined
significance (MGUS) and smoldering multiple myeloma (SMM): novel biological insights and
development of early treatment strategies". Blood (journal) 117 (21): 5573-5581.
[8] Kumaresan PR, Lai WC, Chuang SS, Bennett M, Mathew PA. CSl, a novel member of the CD2
family, is homophilic and regulates NK cell function. Mo! Immunol 2002;39: 1-8.
[9] Hsi ED, Steinle R, Balasa B, et al. Expression of CS 1 (SLAMF7) in benign and neoplastic plasma
cells: a potential new therapeutic target for the treatment of multiple myeloma. Clin Cancer Res
2008;14:2775-84. Abstract/FREE Full Text
[10] Shaughnessy JD Jr., Zhan F, Burington BE, et al. A validated gene expression model of high-risk
multiple myeloma is defined by deregulated expression of genes mapping to chromosome 1. Blood
2007; 109:2276-84.
[11] Hsi ED, Steinle R, Balasa B, et al. CS 1, a potential new therapeutic antibody target for the
treatment of multiple myeloma. Clin Cancer Res 2008;14(9):2775-2784.
[12] Bensinger W, Zonder J, Singhal S, et al. Phase I trial of HuLuc63 in multiple myeloma. Blood
2007;110:358a.
[13] Casucci, Monica; Attilio Bondanza (2011). "Suicide Gene Therapy to Increase the Safety of
Chimeric Antigen Receptor-Redirected T Lymphocytes". Journal of Cancer 2: 378-382.
[14] Pule, M; Finney H; Lawson A (2003). "Artificial T-cell receptors". Cytotherapy 5 (3): 211-26.
[15] Magee MS, Snook AE. Challenges to chimeric antigen receptor (CAR)-T cell therapy for cancer.
Discov Med. 2014 Nov;l8(100):265-71.
[16] Porter DL, Levine BL, Kalos M, Bagg A, June CH. Chimeric antigen receptor-modified T cells in
chronic lymphoid leukemia. N Engl J Med. 2011;365:725-733.
[17] Grupp SA, Kalos M, Barrett D, Aplenc R, Porter DL, Rheingold SR, et al. Chimeric antigen
receptor-modified T cells for acute lymphoid leukemia. N Engl J Med. 2013;368:1509-1518.
[18] Kalos M, Levine BL, Porter DL, Katz S, Grupp SA, Bagg A, et al. T cells with chimeric antigen
receptors have potent antitumor effects and can establish memory in patients with advanced leukemia.
Sci Transl Med. 2011;3:95ra73.
19
[19] Maus MV, June CH. Zoom Zoom: racmg CARs for multiple myeloma. Clin Cancer Res.
2013;19: 1917-1919.
[20] Bae J, Song W, Smith R, Daley J, Tai YT, Anderson KC, et al. A novel immunogenic CS 1-specific
peptide inducing antigen-specific cytotoxic T lymphocytes targeting multiple myeloma. Br J Haematol.
2012;157:687-701.
20
Abstract (if available)
Abstract
Currently, no effective therapies exist for human Primary effusion Lymphoma (PEL). Here we used a humanized CS1 monoclonal antibody (mAb) to target CS1, which is a cell surface glycoprotein that selectively and consistently expressed on B-cell cancer, while PEL is also kind of B-cell lymphoma, to treat PEL. Chimeric antigen receptors (CARs), also known as artificial T cell receptors, are a modification of extracted patient T-cells that improve on human existing recognition abilities. We used CARs as our CS1 mAb carrier which were then transfected to lentiviral vectors and infected NK92-MI cells, which can be treated to PEL cell lines. Administration of in vitro cytotoxicity assay showed significantly inhibition of tumor expression. Our study suggests that anti-CS1 CAR may represent a promising approach for the treatment of PEL.
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Asset Metadata
Creator
Gong, Songjie
(author)
Core Title
Construction and testing of chimeric antigen receptor targeting CS1 for treatment of primary effusion lymphoma
School
Keck School of Medicine
Degree
Master of Science
Degree Program
Molecular Microbiology and Immunology
Publication Date
07/30/2015
Defense Date
06/19/2015
Publisher
University of Southern California
(original),
University of Southern California. Libraries
(digital)
Tag
chimeric antigen receptor,CS1,multiple myeloma,OAI-PMH Harvest,primary effusion lymphoma
Format
application/pdf
(imt)
Language
English
Contributor
Electronically uploaded by the author
(provenance)
Advisor
Chaudhary, Preet M. (
committee chair
), Machida, Keigo (
committee member
), Zandi, Ebrahim (
committee member
)
Creator Email
511531829@qq.com,songjieg@usc.edu
Permanent Link (DOI)
https://doi.org/10.25549/usctheses-c3-615523
Unique identifier
UC11302821
Identifier
etd-GongSongji-3746.pdf (filename),usctheses-c3-615523 (legacy record id)
Legacy Identifier
etd-GongSongji-3746.pdf
Dmrecord
615523
Document Type
Thesis
Format
application/pdf (imt)
Rights
Gong, Songjie
Type
texts
Source
University of Southern California
(contributing entity),
University of Southern California Dissertations and Theses
(collection)
Access Conditions
The author retains rights to his/her dissertation, thesis or other graduate work according to U.S. copyright law. Electronic access is being provided by the USC Libraries in agreement with the a...
Repository Name
University of Southern California Digital Library
Repository Location
USC Digital Library, University of Southern California, University Park Campus MC 2810, 3434 South Grand Avenue, 2nd Floor, Los Angeles, California 90089-2810, USA
Tags
chimeric antigen receptor
CS1
multiple myeloma
primary effusion lymphoma