|
| Benjamin
Storek |
 |
|
Home
Institution
Humboldt-Universität Berlin
Host
Institution(s)
Department of Oncology, Mount Sinai School of Medicine, New York City
Research Mentor: Andreas S. Beutler, M.D.
E-Mail:
b.storek@gmx.de
|
Research
Topic
see Abstract |
Personal
Reactions to the U.S. Experience
The Biomedical Exchange Program (BMEP) made one of my favorite dreams possible:
to live in New York City. It was amazing and I have enjoyed every minute.
NYC… is GREAT! Very interesting and informative lectures, seminars and meetings
at Mount Sinai! There was great concern over my welfare and education from
my mentor as well as other professors! I had never learned so many different
things before! My greatest challenge was my work with rodents. In the beginning,
I needed a lot of time and motivation just to move a rat from one cage to
another (I was soaked in sweat and my hands were shaking tremendously).
NYC… is AMAZING! Soccer games in Central Park, robot parties and Super Bowl
parties! The people in the labs were always very nice and very helpful!
I made fantastic friends in the labs, in the Fifth Avenue Presbyterian Church
I attended, and in the Oratorio Society Choir in which I sang. It was with
them that I had really great times (especially the conversations and discussions)!
One of the greatest experiences was singing in the Oratorio Society Choir
of NYC with the more than 200 people in the choir, however, you must have
a successful audition. The rehearsals were fun and sometimes very difficult,
but we had performances in Carnegie Hall! I never thought I would have the
possibility to sing in this famous hall.
NYC… is BRILLIANT! Concerts, basketball games, baseball games, and Broadway
shows! Of course, tickets are usually quite expensive, but students get
very cheap discounts from their universities.
NYC… is SHOCKING! There are so many homeless people in a society with supposedly
high living standards! There are a lot of rats, especially in subway stations!
And most of the things in the stores are overpriced! |
Greatest
Difficulties Encountered
At the Halloween Parade I was approached by two officers in civilian clothes
because other officers had described someone of my description to be drinking
alcohol on the street. But I did not have an open alcohol container, a bag
or anything else. The officers had a talk with me and did not accept my
friends as witnesses, and also refused to explain the reasons and validity
behind these accusations. In the end, I had two court procedures which I
finally won and my case was dismissed. |
Most
humorous incident
|
|
Helpful
Hints for Future Students
- I recommend
applying for a research program at the MSSM: www.mssm.edu
- If you
are interested in going to a church service in NYC I recommend the Fifth
Avenue Presbyterian Church: www.fapc.org
|
Acknowledgements
Support for these studies is provided by the Biomedical Sciences Exchange
Program (BMEP), the American Society for Clinical Oncology (ASCO), the Martell
Foundation, and the National Institute of Neurological Disorders and Stroke
(NINDS) of the National Institutes of Health (NIH). |
|
|
| Abstract
on Research Topic |
|
Intrathecal
Gene Transfer for Chronic Cancer Pain
|
| Authors:
Benjamin Storek, William G.M. Janssen, Christopher E. Walsh, John H. Morrison,
and Andreas S. Beutler |
Institution:
The Samuel Bronfman Department of Medicine (Oncology) and The Fishberg Center
for Neurobiology, Mount Sinai School of Medicine, New York, NY |
Introduction:
Unrelieved chronic pain from advanced cancer and debilitating side effects
from opioids like morphine (used to relief the pain) are among the most
disruptive events for patients with cancer and their families. In the eye
of patients and the lay public, chronic pain is still closely linked to
advanced cancer. The relief of suffering from pain is seen as a major goal
of medical treatment. Despite significant advances in palliative care, cancer
pain treatment still frequently fails even under optimal circumstances like
well-trained, attentive physicians, ready access to opioid medications,
and use of non-opioid adjuvants. The reason is that systemic opioids like
morphine often induce debilitating side effects, which are as bad as or
worse than the cancer pain that they are expected to ameliorate. The mentor's
laboratory has developed a new approach to pain treatments for cancer patients
based on intrathecal gene transfer. The identification of the optimal gene
transfer vector is one of the major objectives that are currently under
investigation. In this context, the BMEP-supported student is studying gene
transfer to the spinal cord and the surrounding meningeal linings using
a new type of vector, namely a self-complementary variant of recombinant
Adeno-Associated Virus (sc-AAV). The spinal cord is a center for pain control
making it a logical target for the development of new selective treatments. |
Methods:
1. Intrathecal vector delivery: A catheter consisting of pre-stretched PE10
tubing was inserted into the lumbar subarachnoid space in rats through an
opening in the cisterna magna. The virus was delivered (virus dose of 9x109
per animal) and after administration the catheter was removed. On mice the
injections procedure will be involved directing a 30-gauge needle, matched
to a 25 ml syringe, into an intervertebral space at approximately the level
of the fifth of sixth lumbar vertebrae (Hylden and Wilcox et al. 1980).
2. Quantification of EGFP expression by Western blot analysis: For this
technique frozen and grinded spinal cord, including meningeal linings and
dorsal root ganglia will be used. Immunodetection will be performed using
a mouse monoclonal antibody against EGFP (JL-8, Clontech) as a primary antibody
and an anti-mouse antibody coupled to horse reddish peroxidase (Amersham)
as a secondary antibody.
3. Histologic analysis of EGFP expression: To identify the type of transduced
cells we will detect EGFP by fluorescence microscopy. All animals will be
perfused transcardially with cold 2% paraformaldehyde. After perfusion,
the spinal cord, meningeal linings, and dorsal root ganglia will be removed,
postfixed for 6 hours in the same fixative and then immersed in increasing
concentrations of sucrose in PBS (10%, 20%, and 30%). The tissue will be
frozen on dry ice and cut at 50 mm on a freezing microtome. For the detection
of EGFP a polyclonal antibody against EGFP (ab290, Abcam) will be used and
the sections will be processed for fluorescence and analyzed on confocal
laser scanning microscope as above using the Zeiss Plan-Apochromat 16x/1.40-100x/1.40
oil immersion objectives with a digitally fixed aperture. |
Results/Conclusion:
Preliminary data suggests the following key findings:
1. scAAV leads to true long term gene expression in the spinal cord compartment
following intrathecal administration, i.e. by lumbar puncture. We have observed
gene expression up to four months, our longest experimental time point.
2. The choice of scAAV serotyp may be a critical parameter. Our data suggest
that the serotypes 1 and 5 are markedly superior to the serotype 2, which
has been traditionally used for AAV gene transfer. Previous data from the
mentor's laboratory has shown that intrathecal gene transfer is an effective
modality of pain treatment in rodent models. But the duration of effect
was limited due to shortcomings of the gene transfer vector. Our data suggests
that scAAV may resolve these limitations and thus become a viable strategy
for further therapeutic developments. We are currently in the planning stage
for experiments involving large animals (pigs, monkeys). |
|
