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`XIV
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`ll. Molekulare Onkologie 1
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`Die Rolle der Apoptose—regulierenden Proteine Bel—XL und Bax
`in der durch Gemcitabin induzierten Cytotoxizitat beim Pankreaskarzinom
`The role of the apoptosis—regulatingproteins Bcl-xL and Bax
`in gemcitabine—induced cytotoxicity in pancreatic carcinoma
`
`H. Ungefroren, B. Schniewind, D. Henne—Bruns und H. Kalthoff
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`. 25
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`Inhibition von ICE (Caspase—l) induziert Zelltod in Pankreaskarzinomzellen
`Inhibition ofICE (caspase—1) induces cell death in pancreatic carcinoma cells
`
`S. Schlosser, F. Gansauge, M. Ramadani, H. G. Beger und S. Gansauge
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`. 29
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`Prevention von Metastasenwachstum beim Pankreaskarzinom
`
`in vivo durch Apoptoseinduktion mittels Genistein
`Prevention of metastatic pancreatic cancer growth in vivo by induction
`ofapoptosis with genistein
`
`M. W. Miiller, P. Biichler, H. A. Reber, O. J. Hines, H. Friess, M. W. Biich‘ler
`und H. G. Beger
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`1
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`. 33
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`Rapamycin inhibiert das Tumorwachstum und die Tumormetastasierung
`fiber Antiangiogenese
`Rapamycin inhibits tumor growth and metastasis by antiangiogenesis
`
`M. Guba, P. von Breitenbuch, E. Geissler, S. Farkas, C. Ziilke, M. Anthuber,
`K.—W. Iauch und M. Steinbauer
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`. 37
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`Anti-angiogene Gentherapie mittels eines dominant negativen VEGF—RZ
`kodierendem Retroviruses
`
`Antiangiogenic gene therapy with a dominant negative VEGF—RII mutant retrovirus
`
`P. Biichler, H. A. Reber, M. W. B'Lichler, H. Friess, A. Ullrich, O. I. Hines und H. G. Beger
`
`. 41
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`Anti—angiogenetische Therapie zur Behandlung vom humanen Pankreaskarzinom
`nach orthotoper Implantation in die Nacktmaus durch Blockade von NF K‘B
`mit Hilfe des Proteasom Inhibitor PS—341
`
`Antiangiogenic therapyfor the treatment ofhuman pancreatic cancer orthotopically
`implanted in nude mice via the inhibition OfNFICB using the proteasome inhibitor
`PS—341
`
`C. I. Bruns, M. T. Harbsion, R. 1. Bold, I. P. Elliot, I. Adams, I. Abbruzzese,
`A. H. Holscher und D. I. McConkey
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`. 45
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`Ill. Molekulare Onkologie 2
`
`Expression der Multi—Drug—Resistance—Gene mdrl und mrp
`in kolorektalen Karzinomen
`
`Expression of multidrug—resistance genes mdr1 and mrp in colorectal carcinoma
`
`C. G. Schneider, S. B. Hosch, A. Reyrnann, G. Froschle, I.—H. Brasen und I. R. Izbicki
`
`. 49
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`Rapamycin inhibiert das Tumorwachstum
`und die Tumormetastasierung iiber Antiangiogenese
`
`Rapamycin inhibits tumor growth and metastasis by antiangiogenesis
`
`M. Guba1, P. von Breitenbuch2, E. Geissler2
`und M. Steinbauer1
`1 Klinik und Poliklinik fur Chirurgie
`2 Chirurgische Forschung der Universitat Regensburg
`
`, S. Farkas1, C. Ziilke1, M.Anthuber1, K.-W. Jauch1
`
`Abstract
`
`Conventional immunosuppressive drugs have been used effectively to prevent immuno(cid:173)
`logic rejection in organ transplantation. However, cancer development and recurrence are
`ominous risk factors for these immunocompromised patients. In the present study we
`show that the new immunosuppressive drug rapamycin may have a unique ability to re(cid:173)
`duce the risk of cancer development, while simultaneously providing effective immuno(cid:173)
`suppression. Experimentally, rapamycin inhibited tumor metastasis and angiogenesis in
`in vivo mouse models. Furthermore, rapamycin demonstrated antiangiogenic activities
`linked to vascular endothelial growth factor antagonism. In contrast, the most widely rec(cid:173)
`ognized immunosuppressive drug, cyclosporine, promoted both primary tumor growth
`and metastasis in our models. This study suggests that the use of rapamycin, instead of
`cydosporine, may reduce the chance of recurrent, or de novo, cancer development in
`high-risk transplant patients.
`
`Einleitung
`
`Tumorrezidive nach Transplantation bei Tumorpatienten oder de nova Entstehung von
`malignen Tumoren unter Immunsuppression stellen haufige und gefiirchtete Komplika(cid:173)
`tionen dar. Die Immunsuppression an·sich erhoht das Risiko einer de novo Malignom(cid:173)
`entstehung um das 3-4-fache [3]. 54% der Patienten, die nach einer erfolgreichen Tu(cid:173)
`mortherapie aufgrund einer Nierentransplantation immunsuppremiert werden, erfoiden
`ein Tumorrezidiv innerhalb von 2 Jahren [1]. Nach Lebertransplantation aufgrund eines
`Cholangiokarzinoms wurden mit 51 % etwa vergleichbar hohe Rezidivraten festgestellt
`[2]. Daher stellen Malignome nach erfolgreicher Transplantation, die haufigste Todesur(cid:173)
`sache bei immunsuppremierten Patienten dar. Um die Malignomrate bei diesem Patien(cid:173)
`tengut zu verringern, sollten optimaler Weise Immunsuppressiva verwendet werden, die
`gleichzeitig eine antitumorale Potenz besitzen. In der vorliegenden Arbeit zeigen wir, dass
`der Einsatz von Rapamycin als Immunsuppressivum diesem Anforderungsprofil entspre(cid:173)
`chen konnte.
`
`Chirurgisches Forum 2001, Band 30
`
`.998) Bax, but
`
`human breast
`
`~olen J, Myers
`ociated Tyro-
`
`rersi ta tskli(cid:173)
`:hael.muel-
`
`:f
`
`,;I
`
`. I
`
`I
`
`.. I
`; I
`
`LJ1
`
`I
`I
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`38
`
`Methodik
`
`Der Einfluss einer immunsuppressiven Dosis von Cyclosporin (10 mg/kg i.p. tagl.) und
`Rapamycin (1,5 mg/kg i.p. tagl.) im Vergleich zur Kontrollgruppe (Vehikel) auf die Leber- .
`metastasierung wurde am Tag 10 nach intraportaler Injektion von 3 x io5 CT-26 Maus
`Kolonkarzinomzellen bestimmt. Neben der Bestimmung des Lebergewichts und der
`Oberflachenmetastasierung wurden die Lebern histologisch untersucht. Der Serum vas(cid:173)
`cular endothelial growth factor (VEGF) Spiegel der Tiere wurde mittels ELISA gemessen
`(n = 7). In der transparenten Ri.i~kenhautkammer wurde zusatzlich der Einfluss der Im(cid:173)
`munsuppressiva auf das Tumorwachstum [Tumorvolumen (TV)] und die Tumorneoan(cid:173)
`giogenese [Mikrovaskulare Dichte (MVD)] unter zu Hilfenahme der Intravitalmikrosko(cid:173)
`pie am Tag i, 3, 5, 7, 9 und 11 bestimmt (n = 7 ). *p < 0,05 vs. Kontrolle.
`
`Ergebnisse
`
`Die tagliche Gabe von Rapamycin in immunsuppressiver Dosierung (Talspiegel am Tag
`10: Rapamycin 39 ± 6 ng/ml vs. Cyclosporin 118 ± 21 ng/ml) fiihrte zu einer deutlichen Re(cid:173)
`duktion der Metastasierung von .CT-26 Zellen. Im Gegensatz zu Cyclosporin und der Kon(cid:173)
`trollgruppe zeigten Rapamycin behandelte Tiere ein signifikant geringeres Lebergewicht
`(Cyclosporin 1,9 ± 0,1* vs. Kontrolle 1,7 ± 0,1 vs. Rapamycin 1,3 ± 0,1* g) sowie eine signi(cid:173)
`fikante Reduktion der Anzahl von Oberflachenmetastasen (Cyclosporin 203 ± 32* vs.
`Kontrolle 155 ± 32 vs. Rapamycin 25 ± io*) am Tag 10 nach intraportaler Injektion der Tu(cid:173)
`morzellen. Die histologische Auswertung der Leberschnitte zeigte bei Cyclosporin be(cid:173)
`handelten Tieren invasiv wachsende, gefaBreiche Tumore, wahrend Rapamycin behan(cid:173)
`delte Tiere zum gleichen Zeitpunkt kleine, disseminierte und auffallend avaskulare Tu(cid:173)
`more aufwiesen. Rapamycin fiihrte dabei zu einer signifikanten Verringerung der Serum
`VEGF Spiegel (Cyclosporin 79 ± 14* vs. Kontrolle 60 ± 3 vs. Rapamycin 38 ± 2* pg/ml).
`Dariiberhinaus fiihrte Rapamycin im Vergleich zur Kontrollgruppe zu einem signifikant
`verringerten Tumorwachstum (TV am Tag i3: Cyclosporin 207 ± Bo* vs. Kontrolle
`i58 ± 56 vs. Rapamycin 33 ± i2* mm3) in der transparenten Ruckenhautkammer. In der
`Intravitalmikroskopie konnten wir eine deutliche Verminderung der Tumorneoangioge(cid:173)
`nese (MVD am Tag 11: Cyclosporin 179 ± 14 * vs. Kontrolle 1~6 ± 9 vs. Rapamycin 35 ± 6*
`cm- 1
`) als Ursache fiir dieses Phanomen identifizieren. Die Immunsuppression mit Cyclo(cid:173)
`sporin zeigte bei beiden Parametern den gegenteiligen Effekt.
`
`Diskussion und Schlussfolgerung
`
`Die vorliegenden Daten im Mausmodell zeigen, class Rapamycin in einer immunsuppres(cid:173)
`siven Dosierung eine zusatzliche antitumorale Wirkung besitzt. Dieser Effekt auf das Tu(cid:173)
`morwachstum beruht auf einer antiangiogenetischen Wirkung, die auf eine verringerte
`VEGF Produktion der Tumorzellen zuriickgefiihrt werden konnte. Diesen Ergebnissen
`kommt eine besondere Bedeutung zu, da zum gegenwartig~n Zeitpunkt davon ausgegan(cid:173)
`gen werden muss, dass die bisherige Immunsuppression sowohl die de novo Tumorent(cid:173)
`stehung, als auch die Tumorprogression offensichtlich begilnstigt [4J. Aus diesem Grund
`halten wir die Uberprilfung dieser experimentellen Daten in kontrollierten Studien an
`
`Patienten, die
`nome transpl
`sporin einges
`
`Literatur
`
`i. Penn I (2000
`2. MeyerCG,Pe
`Trans plan ta ti
`3. Penn I. (1998
`147-158
`4. Hojo M, Mor
`Cyclosporine
`
`Korresponde1
`sitat Regenst
`6801, Fax: +-t
`
`f
`' I
`~
`. l
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`Breckenridge Exhibit 1061
`Guba
`Page 004
`
`

`

`Patienten, die wegen einer Tumoranamnese oder bei erhohtem Risiko fiir de nova Karzi(cid:173)
`nome transplantiert werden, fiir notwendig. Dabei sollte Rapamycin alternativ zu Cyclo(cid:173)
`sporin eingesetzt werden.
`
`39
`
`Literatur
`
`1. Penn I (2000) Cancers in renal transplant recipients.Adv Ren Replace Ther 7: 147-156
`2. Meyer CG, Penn I, James L (2000) Liver transplantation for cholangio-carcinoma: results in 207 patients.
`Transplantation 69: 1633 -1637
`.
`3. Penn I. (1998) Occurrence of cancers in immunosuppressed organ transplant recipients. Clin Transpl
`'147-158
`4. Hojo M, Morimoto T, Maluccio M, Asano T, Morimoto K, Lagman M, Shimbo T, Suthanthiran M (1999)
`Cyclosporine induces cancer progression by a cell-autonomous mechanism. Nature 397: 530 - 534
`
`Korrespondenzadresse: Dr. med. M. Guba, Klinik und Poliklinik fiir Chirurgie der Univer(cid:173)
`sitat Regensburg, Franz-Josef-StrattB-Allee 11, 93042 Regensburg, Tel.: ++49(0) 941/944-
`6801, Fax: ++49(0) 941/944-6.802, e-mail: markus.guba@klinik.uni-regensburg.de
`
`p. tagl.) und
`lf die Leber(cid:173)
`CT-26 Maus
`its und der
`·Serum vas(cid:173)
`;A gemessen
`luss der Im-
`1morneoan(cid:173)
`almikrosko-
`
`egel am Tag
`utlichen Re(cid:173)
`nd der Kon(cid:173)
`ebergewicht
`~ eine signi-
`03 ± 32* vs.
`tion der Tu(cid:173)
`:>sporin he(cid:173)
`rein behan(cid:173)
`skulare Tu(cid:173)
`~ der Serum
`: 2* pg/ml).
`signifikant
`:. Kontrolle
`mer. In der
`.eoangioge(cid:173)
`rcin 35 ± 6*
`. mit Cyclo-
`
`msuppres(cid:173)
`mf <las Tu(cid:173)
`rerringerte
`rgebnissen
`ausgegan(cid:173)
`fumorent(cid:173)
`em Grund
`:tudien an
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`Guba
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`
`

`

`Rapamycin inhibits tumor growth and metastasis by
`antiangiogenesis
`
`
`
`
`
`M. Guba1, P. von Breitenbuch2, E. Geissler2, S. Farkas1, C. ZüIke1, M. Anthuber1, K.-
`W. Jauch1 and M. Steinbauer1
`
`1 Surgical Clinic and Polyclinic
`
`2 Surgical Research at the University of Regensburg
`
`
`
`
`
`Abstract
`
`Tumor recurrence after transplantation in tumor patients or de novo malignant tumor
`development in patients whilst immunosuppressed constitute a frequent and dreaded
`complication. Immunosuppression increases the risk of de novo malignant tumor
`development 3 to 4 fold [3]. A total of 54% of patients who are immunocompromised
`due to a kidney transplant following successful tumor therapy suffer from tumor
`recurrence within 2 years [1]. Following liver transplants due to cholangiocarcinoma, a
`comparatively high recurrence rate of 51% was observed [2]. This makes malignant
`tumors after successful
`transplantation
`the most
`frequent cause of death
`in
`immunocompromised patients. To lower the malignant tumor rate in this patient
`population, ideally immunosuppressive drugs should be used which also exhibit
`antitumor potency. In the present paper, we demonstrate that the use of rapamycin as
`an immunosuppressive drug could meet these requirements.
`
`-----------------------------------------------
`
`Surgical Forum 2001, Volume 30
`
`
`
`
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`Guba
`Page 006
`
`

`

`
`
`Methodology
`
`The influence of an immunosuppressive dosage of cyclosporine (10 mg/kg i.p. daily)
`and rapamycin (1.5 mg/kg i.p. daily), compared to the control group (excipient), on liver
`metastasis was determined on day 10 after the intraportal injection of 3 x 105 CT-26
`mouse colon carcinoma cells. In addition to determining the weight of the liver and
`surface metastasis, the livers were histologically analyzed. The serum vascular
`endothelial growth factor (VEGF) levels of the animals were measured using ELISA (n
`= 7).
`In
`the
`transparent skinfold chamber, additionally
`the
`influence of
`the
`immunosuppressive drugs on tumor growth [tumor volume (TV)] and the tumor
`neoangiogenesis [microvascular density [MVD)] was determined using intravital
`microscopy on days 1, 3, 5, 7, 9 and 11 (n = 7). *p < 0.05 vs. control group.
`
`Results
`
`The daily administration of an immunosuppressive dosage of rapamycin (trough level
`on day 10: rapamycin 39 ± 6 ng/ml vs. cyclosporine 118 ± 21 ng/ml) resulted in a
`considerable decrease in CT-26 cell metastasis. In contrast to cyclosporine and the
`control group, animals treated with rapamycin exhibited a significantly lower liver
`weight (cyclosporine 1.9 ± 0.1* vs. controls 1.7 ± 0.1 vs. rapamycin 1.3 ± 0.1* g), and
`the number of surface metastases was significantly reduced (cyclosporine 203 ± 32*
`vs. controls 155 ± 32 vs. rapamycin 25 ± 10*) on day 10 after the intraportal injection of
`the tumor cells. The histological evaluation of the liver sections showed invasive
`growth of tumors containing plenty of vessels in animals treated with cyclosporine,
`while animals treated with rapamycin contained small, disseminated and strikingly
`avascular tumors over the same time frame. Rapamycin resulted in a significant
`reduction in the serum VEGF level (cyclosporine 79 ± 14* vs. controls 60 ± 3 vs.
`rapamycin 38 ± 2* pg/mI). Moreover, compared to the control group, rapamycin
`resulted in significantly reduced tumor growth (TV on day 13: cyclosporine 207 ± 80*
`vs. controls 158 ± 56 vs. rapamycin 33 ± 12* mm3) in the transparent skinfold
`chamber. In intravital microscopy we were able to identify a considerable decrease in
`tumor neoangiogenesis (MVD on day 11: cyclosporine 179 ± 14* vs. controls 116 ± 9
`vs. rapamycin 35 ± 6* cm-1) as the cause for this phenomenon. Immunosuppression
`with cyclosporine showed the opposite effect for both parameters.
`
`Discussion and conclusion
`
`The present data from the mouse model shows that rapamycin, when used in an
`immunosuppressive dosage, has an additional antitumor effect. This effect on tumor
`growth is based on antiangiogenetic action, which we were able to attribute to lower
`VEGF production of the tumor cells. These results are particularly important since it
`must be assumed at the present time that immunosuppression seems to favor both de
`novo tumor development and tumor progression [4]. For this reason, we consider it
`necessary to review this experimental data in controlled studies on patients who
`
`
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`Guba
`Page 007
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`

`

`receive transplants due to tumor anamnesis or an increased risk for de novo cancer.
`Rapamycin should be used as an alternative to cyclosporine.
`
`
`
`
`
`Literature
`
`1. Penn I (2000) Cancers in renal transplant recipients. Adv Ren Replace Ther 7:147-156
`
`2. Meyer CG, Penn I, James L (2000) Liver transplantation for cholangio-carcinoma: results in 207
`patients. Transplantation 69: 1633 -1637
`
`3. Penn I. (1998) Occurrence of cancers in immunosuppressed organ transplant recipients. Clin Transpl
`147-158
`
`4. Hojo M, Morimoto T, Maluccio M, Asano T, Morimoto K, Lagman M, Shimbo T, Suthanthiran M (1999)
`Cyclosporine induces cancer progression by a cell-autonomous mechanism. Nature 397: 530-534
`
`
`
`Correspondence address: Dr. med. M. Guba, Surgical Clinic and Polyclinic of the
`University of Regensburg, Franz-Josef-Strauß-Allee 11, 93042 Regensburg, Tel.:
`++49(0) 941/944-6801, Fax: ++49(0) 941/944-6802, E-mail: markus.guba@klinik.uni-
`regensburg.de
`
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`Guba
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`
`

`

`May 18,2017
`
`Certification
`
`Park IP Translations
`
`TRANSLATOR'S DECLARATION:
`
`I, Kerstin Roland, hereby declare:
`
`That I possess advanced knowledge of the German and English languages. The
`attached German into English translation has been translated by me and to the
`best of my knowledge and belief, it is a true and accurate translation of:
`Rapamycin inhibiert das Tumorwachstum und die Tumormetastasierung iiber
`Antia ngiogenese
`
`Kerstin Roland
`
`Project Number: MEGOP _.1705_006
`
`15 W. 37th Street 8th Floor
`
`New York, NY 10018
`
`212.581.8870
`
`ParklP.com
`
`
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`Guba
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