Influence of Selectin Mediated Rolling/Adhesion Dynamics in Prostate Cancer Metastasis

182 Pages·2014·2.02 MB·English

by  Sayeda Karim

Influence of Selectin Mediated Rolling/Adhesion Dynamics in Prostate Cancer
Metastasis: a Micro-Channel Model

By

Sayeda Yasmin-Karim

Submitted in Partial Fulfillment of the Requirement for the Degree
Doctor of Philosophy

Supervised by
Professor Yi-Fen Lee
Professor David Wu

Department Of Chemical Engineering
Arts, Sciences and Engineering
Edmund A. Hajim School of Engineering and Applied Sciences

University of Rochester
Rochester, New York
2014
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Dedication

To my parents: Setara Rahman (mom) and Sadequr Rahman (dad)
My husband: Khondkar Ehsanul Karim; daughter: Moureen Farnaz Karim; and
son: Mahvir Pashin Karim
For their love, support and silent sacrifice
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Biographical Sketch

Sayeda Yasmin-Karim was born in Comilla, Bangladesh. She received her degree in
Medicine (MBBS/MD) from Chittagong Medical College, Chittagong, Bangladesh in 1988. She
then worked as Medical Officer for the Institute of Post Graduate Research Hospital, Dhaka,
Bangladesh from 1989 -1991. She continued to work as Medical Officer for the Health and
Family Planning Department in Bangladesh from 1991 to 1994. She attended Long Island
University, C.W. Post Campus, New York from 1998 to 2001, and graduated with Master of
Science in Medical Biology (Major in Immunology and Minor in Hematology). She worked as
Research Associate in the Department of Biochemistry and Biophysics, School of Nursing, and
Department of Biomedical Engineering at the University of Rochester from 2001 to 2008.
In 2008 she joined the PhD Program in the Department of Chemical Engineering at the
University of Rochester. She pursued her research in prostate cancer under the guidance of Dr.
Yi-Fen Lee. She was granted a Predoctoral Training Award from U.S. Department of Defense
Prostate Cancer Research Program in 2011.
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List of Publications
1. Sayeda Yasmin-Karim, Michael R. King, Edward Messing and Yi-Fen Lee. E-seletin
Ligand-1 Controls Circulating Prostate Cancer Cell Rolling/Adhesion and Metastasis.
Under review.

2. Sayeda Yasmin-Karim, and Yi-Fen Lee. Role of Circulatory Tumor Cell to endothelial
adhesion molecules in solid tumor metastasis process. Manuscript under preparation.

3. Sayeda Yasmin-Karim, Peng-Nien Yin, Christopher R. Silver, Chia-Hao Wu and Yi-
Fen Lee. VDR Plays Role in CXCR-1 and IL-6 cytokine mediated conversion of M2
macrophage during prostate cancer progression. Manuscript under preparation.

4. Huei-Ju Ting, James Messing, Sayeda Yasmin-Karim and Yi-Fen Lee. Identification of
microRNA-98 as a therapeutic target inhibiting prostate cancer growth and a biomarker
induced by vitamin D. The Journal of Biological Chemistry, January 4, 2013 288, 1-9.

5. Huei-Ju Ting, Sayeda Yasmin-Karim, Shian-Jang Yan, Jong-Wei Hsu, Tzu-Hua Lin,
Weisi Zeng, James Messing, Tzong-Jeng Sheu, Bo-Ying Bao, Willis X. Li Edward
Messing and Yi-Fen Lee. A Positive Feedback Signaling Loop between ATM and the
Vitamin D Receptor Is Critical for Cancer Chemoprevention by Vitamin D. Cancer
Research. February 15, 2012 72; 958.

6. SayedaYasmin-Karim, Huei-Ju Ting, Michael King, Edward Messinge and Yi-Fen
Lee. Capture and Analyzing of Prostate Cancer Stem cells using a Microcirculatory
Channel. Proceeding of the American Urological Association (AUA) 2012 Annual
Scientific Meeting.

7. Jong-Wei Hsu, Sayeda Yasmin-Karim, Michael R. King, Joel C. Wojciechowski,
Deanne Mickelsen, Martha L. Blair, Huei-Ju Ting, Wen-Lung Ma, Yi-Fen Lee.
Suppression of Prostate Cancer Cell Rolling and Adhesion to Endothelium by 1α,25-
Dihydroxyvitamin D. The American Journal of Pathology. Volume 178, Issue 2 , Pages
872-880, February 2011.

8. Bo-Ying Bao, Huei-Ju Ting, Jong-Wei Hsu, Sayeda Yasmin-Karim, Edward Messing,
Yi-Fen Lee. Down-regulation of NFκB signals is involved in loss of 1α25-
dihydroxyvitamin D responsiveness. The Journal of Steroid Biochemistry and Molecular
3
Biology. Volume 120, Issue 1, 1 May 2010, Pages 11–21
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Symposium Presentations

1. Sayeda Yasmin-Karim, Michael R. King, Edward Messing and Yi-Fen Lee. E-seletin
Ligand-1 Controls Circulating Prostate Cancer Cell Rolling/Adhesion and Metastasis.
American Urological Association (AUA), Annual Scientific Meeting, 2014.

2. Sayeda Yasmin-Karim, Huei-Ju Ting, Michael R. King, Edward Messing and Yi-Fen
Lee. Capture and Analyzing of Prostate Cancer Stem cells using a Microcirculatory
Channel. University of Rochester Graduate Research Showcase, University of Rochester,
2013.

3. SayedaYasmin-Karim, Huei-Ju Ting, Michael King, Edward Messing and Yi-Fen Lee.
Capture and Analyzing of Prostate Cancer Stem cells using a Microcirculatory Channel.
American Urological Association (AUA), Annual Scientific Meeting, 2012.

4. Sayeda Yasmin-Karim, Jong-Wei Hsu, Huei-Ju Ting, Michael R. King, and Yi-Fen
Lee. Rolling Prostate Cancer Cells in Circulation Potentiate Metastasis: A Microchannel
Model. 15th Annual Scientific Symposium, James P. Wilmot Cancer Center, University
of Rochester, 2010.

5. Sayeda Yasmin- Karim, Michael R. King, and Yi-Fen Lee. Rolling Prostate Cancer
Cells in Circulation Potentiate Metastasis: A Micro-Channel Model. Students Seminar,
Department of Chemical Engineering, HAJIM School of Engineering and Applied
Science, University of Rochester, 2010.
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Acknowledgements

First of all, I would like to express my gratitude to my advisor Dr. Yi-Fen Lee, for giving
me the opportunity to pursue my Ph.D. study. I am so fortunate that she allowed me to join her
group and continued my work for my degree at a critical point in my career. She has always been
incredibly generous with her time, and offered me patience, faith and guidance. Without her
direction and advice I would not have reached this goal. I am also grateful for her endless
assistance in writing my journal article, grant proposal, and thesis. I am deeply appreciative of
my co-advisor from the Chemical Engineering Department, Dr. David Wu, and the members of
my thesis committee Dr. Edward M. Messing and Dr. David G. Foster, for their guidance and
advice in my research. I also thank Dr. Michael R. King, Department of Biomedical Engineering
at Cornell University. The training and guidance I received from his lab when I worked there
really inspired me to pursue my PhD study. Next, I would like to thank Dr. Martha Blair,
Department of Pharmacology and Physiology, for her kindness in allowing me to use her lab at
the beginning of this project when we did not have the facilities, without which this project
would not have seen the light of day. I also wish to express my gratitude to Dr. Hiroshi
Miyamoto, Department of Pathology, Johns Hopkins University School of Medicine, for his
expert opinion regarding the histological slides, which gave me confidence and enthusiasm
during this study. I would also like to thank Dr. Hong Yang, Department of Chemical and
Biomolecular Engineering, University of Illinois at Urbana-Champaign, for his support at the
beginning of my graduate study.
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I would like to express my sincere gratitude to former members of the Lee lab, Dr. Huei-
Ju Ting and Dr. Jong-Wei Hsu (Gus), and to the current lab members Christopher R. Silvers,
Peng-Nien Yin (Benny) and Chia-Hao Wu (Bean) for their scientific insights and technical
expertise. I am also grateful to Patricia Conway for her support and expertise in editing my
thesis, my journal article, and my successful grant proposal. I would like to thank Susan Schoen
for her assistance as lab manager. I also want to express my gratitude to Gina Eagan, Sandra
Willison and Victoria Heberling of the Department of Chemical Engineering at the University of
Rochester for their unfailing assistance throughout my graduate study.
Finally, I have to thank my husband, Dr. Khondkar Karim, for his understanding,
patience, encouragement and love throughout my Ph.D. studies, and my entire family for their
unconditional love and support. The achievement, if any, I have today is dedicated to them.
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Abstract

Metastasis is the cause of death in most patients with prostate cancer (PCa), and the
second most common cancer in men in America (Lee 2009, Thompson 2013). Circulating PCa
cells preferentially roll and adhere on bone marrow vascular endothelial cells, where abundant E-
selectin and stromal cell-derived factor 1 (SDF-1) are expressed, subsequently initiating a
cascade of activation events that eventually lead to the development of metastases. To elucidate
the importance of circulating PCa cells’ rolling and adhesion behaviors in the development of
metastasis, we applied a dynamic cylindrical flow-based microchannel device that is coated with
E-selectin and SDF-1, mimicking capillary endothelium, allowing PCa cells to roll and adhere
under flow.
We reported that PCa cells’ rolling ability is positively correlated with their
aggressiveness using three pairs of PCa cell lines. Using this device we were able to capture a
small fraction of rolling PCa cells and characterize their behavior and genetic contents. The
rolling cells make up less than 5% of the circulatory tumor cell population. They possess
adhesion ability and display aggressive cancer phenotype with stem like characteristics and
develop metastasis in vivo while the majority of non-rolling/floating cells do not. Moreover, the
expression level of genes associated with cancer stem cells and cancer metastasis was found to
be elevated in rolling PCa cells than those in floating cells. Importantly, the expression of E-
selectin ligans-1(ESL-1) was overexpressed in rolling PCa cells and knock down of ESL-
1significantly impaired PCa cells’ rolling capacity and cancer aggressive phenotypes. Detailed
mechanism studies revealed that ESL-1 activates RAS, consequently inducing a signal cascade
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such as MAP kinase and its downstream targets to influence cell proliferation and migration. To
evaluate further the status of ESL-1 in human PCa samples, we analyzed the gene expression
profile of ESL-1 through public microarray repositories from NCBI Gene Expression Omnibus.
We found elevated levels of ESL-1in metastatic PCa in comparison to normal prostate and
primary tumors, and no significant difference in normal prostate and primary PCa tumors. This
elevated level of ESL-1 found in human PCa metastatic samples further supports ESL-1’s roles
in PCa metastasis.
In summary, we conclude that adhesion molecules play a major role in cancer metastatic
process and this dynamic microchannel system allows us to capture the rolling CTCs and
identify the specific gene profile for each individual cancer. Our results indicated that circulating
PCa cells’ rolling capacity contributes to PCa metastasis, and this is in part controlled by ESL-1
and downstream targeted signals. Thus, ESL-1 and its induced oncogenic pathways represent
new therapeutic targets that control circulating PCa cells’ rolling, adhesion and distant
metastasis. The status of this molecule in circulating PCa cells could also be an important
functional biomarker for prediction of disease behavior. A customized medicine for individual
patients can also be developed using this technique.
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Contributors and Funding Sources

This work was supervised by a dissertation committee consisting of Dr. Yi-Fen Lee (advisor) of
Department of Urology, Dr. David Wu (co-advisor) and Dr. David G. Foster of Department of
Chemical Engineering, and Dr. Edward M. Messing of Department of Urology. It was also
guided by Dr. Michael R. King, Department of Biomedical Engineering at Cornell University.
All work for the dissertation was completed independently by the student, Sayeda Yasmin-
Karim. This study was supported by the Department of Defense Prostate Cancer Research
Program Predoctoral Training Award (# W81XWH-11-1-0429) to Sayeda Yasmin-Karim.