Antibodies directed against the 37 kDa/67 kDa laminin receptor as therapeutic tools for the treatment of prion diseases and cancer [Elektronische Ressource] / Chantal Zuber

Chantal Zuber

Sujets

• Gesundheit

Informations

• Publié par: ludwig-maximilians-universitat_munchen
• Publié le: 01 janvier 2008
• Nombre de lectures: 26
• Langue: English
• Poids de l'ouvrage: 25 Mo

Extrait

Dissertation
zur Erlangung des Doktorgrades
der Fakultät für Chemie und Pharmazie
der Ludwig-Maximilians-Universität München



Antibodies directed against
the 37 kDa/67 kDa laminin receptor
as therapeutic tools for the treatment of
prion diseases and cancer


Chantal Zuber
aus
München
2008
Erklärung

Diese Dissertation wurde im Sinne von §13 Abs. 3 der Promotionsordnung vom 29. Januar
1998 von PD Dr. Stefan Weiss betreut.




Ehrenwörtliche Versicherung
Diese Dissertation wurde selbständig, ohne unerlaubte Hilfe erarbeitet.

München, am 19.06.2008

____________________
Chantal Zuber



Dissertation eingereicht am 19. Juni 2008


1. Gutachter: PD Dr. Stefan Weiß
2. Gutachter: Prof. Dr. Hans A. Kretzschmar


Mündliche Promotionsprüfung am 16. Juli 2008 Im Verlauf dieser Arbeit wurden folgende Originalpublikationen veröffentlicht bzw.
eingereicht:


Zuber, C., Mitteregger, G., Pace., C., Kretzschmar, H. A., Zerr, I. and Weiss, S., Anti-
LRP/LR antibody W3 hampers peripheral PrPSc propagation in scrapie infected mice. Prion
2007. 1: 207-212.


Zuber, C., Knackmuss, S., Rey, C., Reusch, U., Rottgen, P., Frohlich, T., Arnold, G. J., Pace,
C., Mitteregger, G., Kretzschmar, H. A., Little, M. and Weiss, S., Single chain Fv antibodies
directed against the 37 kDa/67 kDa laminin receptor as therapeutic tools in prion diseases.
Mol Immunol 2008. 45: 144-151.


Zuber, C., Mitteregger, G., Schuhmann, N., Rey, C., Knackmuss, S., Rupprecht, W., Reusch,
U., Pace, C., Little, M., Kretzschmar, H. A., Hallek, M., Büning, H. and Weiss, S., Delivery
of single-chain antibodies scFvs directed against the 37 kDa/67 kDa laminin receptor into
mice via recombinant Adeno-associated viral vectors for prion disease gene therapy. J. Gen.
Virol. in press.


Kolodziejczak, D., Zuber, C., Beck, J., Richt, J., Brenig, B., Vana, K. and Weiss, S., CWD
and sheep Scrapie prions colocalize with the 37kDa/67kDa laminin receptor on human
enterocytes. EMBO rep. submitted.


Zuber, C., Knackmuss, S., Zemora, G., Reusch, U., Vlasova, E., Diehl, D., Mick, V.,
Hoffmann, K., Nikles, D., Frohlich, T., Arnold, G. J., Brenig, B., Wolf, E., Lahm, H., Little,
M. and Weiss, S., Invasion of tumorigenic HT1080 cells is impeded by blocking or
downregulating the 37-kDa/67-kDa laminin receptor. J Mol Biol 2008. 378: 530-539.


Die vorliegende Arbeit wurde in der Zeit von November 2005 bis April 2008 im Labor von
PD Dr. Stefan Weiß am Genzentrum der Ludwig-Maximilians-Universität München
angefertigt.


Im Verlauf wurden folgende Übersichtsartikel publiziert:

Rey, C., Zuber, C. and Weiss, S., Therapeutische Ansätze zur Behandlung von
Prionenerkrankungen. Nova Acta Leopoldina 2006. NF 94: 129-144.


Vana, K., Zuber, C., Nikles, D. and Weiss, S., Novel aspects of prions, their receptor
molecules, and innovative approaches for TSE therapy. Cell Mol Neurobiol 2007. 27: 107-
128.


Zuber, C., Ludewigs, H. and Weiss, S., Therapeutic approaches targeting the prion receptor
LRP/LR. Vet Microbiol 2007. 123: 387-393.


Ludewigs, H., Zuber, C., Vana, K., Nikles, D., Zerr, I. and Weiss, S., Therapeutic approaches
for prion disorders. Expert Rev Anti Infect Ther 2007. 5: 613-630.


Vana, K., Zuber, C., Pflanz, H., Kolodiejcak, D., Zemora, G., Bergmann, A-K. and Weiss,
S., LRP/LR as an alternative promising target in therapy of prion diseases. Infectious
Disorders - Drug targets in press.


Vorträge auf Konferenzen
- International Prion Conference, Prion 2007, 26. September 2007, Edinburgh, UK,
Students day, „The laminin receptor as therapeutic target in prion disease“
- International Prion Conference, Prion 2008, 8. – 10. Oktober, 2008, Madrid, Spanien
„Antibodies directed against the prion protein receptor LRP/LR provide alternative
tools in prion diseases“
- NoE Neuroprion Meeting, 28. September 2007, Edinburgh, UK
- NoE Neuroprion Meeting, 14. März 2008, München

ACKNOWLEDGE MENT

I would like to thank PD Dr. Stefan Weiß for giving me the opportunity to intensively study prion
diseases and cancer, visit international prion conferences and giving oral presentations. Furthermore, I
am thankful for the support to fastly proceed my phD. Finally, I would like to thank him for the
creation of the first certificate.

I thank Prof. Dr. Hans A. Kretzschmar for continuous support and for the preparation of the second
certificate.

I would like to thank Prof. Dr. Patrick Cramer, Prof. Dr. Roland Beckmann, Prof. Dr. Dietmar Martin
and Prof Dr. Ralf Jansen for being members of my thesis committee.

I thank Dr. Gerda Mitteregger for continuous support and helpful advice in the animal facility.

I want to thank all present members of the Weiß group, namely Dr. Karen Vana for correcting my
PhD manuscript, for her excellent knowledge, cordiality and high spirits. A big thanks goes to Heike
Ludewigs, sorry, Pflanz, providing everytime a great helping hand not only for the animal facility, but
also for discussing problems and giving me great pharmaceutical advisory (and medication) and
having nice breaks with tasty donats and coffee (Kaffee?).
I´d like to thank Dr. Daphne Nikles for having every time a new idea for going on and giving great
mental help.
A thank goes also to Dr. Clémence Rey, who contributed a lot to my phD.

I thank Katja Vlasova for the contribution of the invasion assays.
I would like to thank Georgeta Zemora, Dominika Kolodziejczak, Cilli Sedlaczek, Katharina
Häussermann, André Heuer and Sonja Perdoch for a really funny and unique working atmosphere.

I want to thank Katharina Krüger and Jennifer Hentrich for the help in producing antibodies and Tina
Hallas for attending me in the animal facility.

Many thanks to Julia Kneißl, for the nice cinema events (every time a good nose for a profitable film!)
and the discussions about research carreer.
A big thank to my boyfriend Marco Niecke for the excellent music support giving me high spirits. I
thank him for the weekend activities searching for sunny places, jogging, swimming and poker.
Finally, I want to thank my parents for supporting me all the time and having always confidence in my
desicions. SUMMARY 3

CHAPTER I 4

Introduction 4
1. Transmissible spongiform encephalopathies 5
2. The cellular prion protein 12
3. The 37 kDa/67 kDa Laminin receptor (LRP/LR) 37
4. Antibodies 47
References 57

Chapter II
Therapeutische Ansätze zur Behandlung von Prionerkrankungen

Chapter III
Novel aspects of prions, their receptor molecules, and innovative approaches
for TSE therapy

Chapter IV
Therapeutic approaches targeting the prion receptor LRP/LR

Chapter V
Therapeutic approaches for prion disorders

Chapter VI
LRP/LR as an alternative promising target in therapy of prion diseases

Chapter VII
Single chain Fv antibodies directed against the 37 kDa/67 kDa laminin
receptor as therapeutic tools in prion diseases


1 Chapter VIII
ScAnti-LRP/LR antibody W3 hampers peripheral PrP propagation in scrapie
infected mice

Chapter IX
CWD and sheep Scrapie prions colocalize with the 37 kDa/67 kDa laminin
receptor on human enterocytes

Chapter X
Invasion of tumorigenic HT1080 cells is impeded by blocking or
downregulating the 37-kDa/67-kDa laminin receptor
ABBREVIATIONS
CURRICULUM VITAE
2 SUMMARY

Transmissible spongiform encephalopathies (TSE), also known as prion diseases, comprise a class
of neurological lethal disorders, affecting both humans and animals. TSEs are commonly associated
with the formation of abnormal protein aggregates and plaques observed in the central nervous
system (CNS) of the affected individual leading to neuronal cell death. The observed aggregates
Scoriginate from accumulation of the abnormal isoform (PrP ) of the host encoded cellular prion
cprotein (PrP ) mainly responsible for the pathology of the disease. Several functions have been
c cdescribed for PrP , however, little is known about the exact role of PrP in the disease and the
underlying mechanisms of accumulation and neurodegeneration, respectively.
The 37 kDa/67 kDa laminin receptor LRP/LR has been identified as a cellular surface receptor for
c Scboth PrP and PrP . In addition, LRP/LR is thought to participate in the prion protein uptake in
intestinal cells such as enterocytes suggesting an implication in the oral prion protein uptake. The
crucial role of LRP/LR in the prion life cycle indicates new possibilities for the development of an
alternative therapy for prion diseases. Tools directed against or downregulating the laminin receptor
have been developed. Among them, anti-LRP/LR antibodies have been proven to block the PrP-
LRP/