Preclinical characterization of the myxobacterial compound pretubulysin as novel vascular disrupting agent:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2013
|
| Schlagworte: | |
| Online-Zugang: | kostenfrei http://d-nb.info/1046503227/34 kostenfrei Inhaltsverzeichnis |
| Beschreibung: | VI, 96 Blatt Ill., graph. Darst. |
| Format: | Langzeitarchivierung gewährleistet, LZA |
Internformat
MARC
| LEADER | 00000nam a2200000zc 4500 | ||
|---|---|---|---|
| 001 | BV041733356 | ||
| 003 | DE-604 | ||
| 005 | 20140327 | ||
| 007 | t | ||
| 008 | 140313s2013 gw ad|| m||| 00||| eng d | ||
| 015 | |a 14,O02 |2 dnb | ||
| 016 | 7 | |a 1046503227 |2 DE-101 | |
| 035 | |a (OCoLC)876897736 | ||
| 035 | |a (DE-599)DNB1046503227 | ||
| 040 | |a DE-604 |b ger |e rakddb | ||
| 041 | 0 | |a eng | |
| 044 | |a gw |c XA-DE-BY | ||
| 049 | |a DE-12 |a DE-19 |a DE-384 |a DE-473 |a DE-703 |a DE-1051 |a DE-824 |a DE-29 |a DE-91 |a DE-1049 |a DE-92 |a DE-739 |a DE-898 |a DE-355 |a DE-706 |a DE-20 |a DE-1102 | ||
| 084 | |a 610 |2 sdnb | ||
| 100 | 1 | |a Kretzschmann, Verena |e Verfasser |4 aut | |
| 245 | 1 | 0 | |a Preclinical characterization of the myxobacterial compound pretubulysin as novel vascular disrupting agent |c Verena Kretzschmann |
| 264 | 1 | |c 2013 | |
| 300 | |a VI, 96 Blatt |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |a München, Univ., Diss., 2013 | ||
| 538 | |a Langzeitarchivierung gewährleistet, LZA | ||
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 700 | 1 | |a Vollmar, Angelika |4 dgs | |
| 776 | 0 | 8 | |i Erscheint auch als |n Online-Ausgabe |o urn:nbn:de:bvb:19-164317 |
| 856 | 4 | 0 | |u https://nbn-resolving.org/urn:nbn:de:bvb:19-164317 |x Resolving-System |z kostenfrei |3 Volltext |
| 856 | 4 | 0 | |u http://d-nb.info/1046503227/34 |x Langzeitarchivierung Nationalbibliothek |
| 856 | 4 | 0 | |u http://edoc.ub.uni-muenchen.de/16431/ |x Verlag |z kostenfrei |
| 856 | 4 | 2 | |m DNB Datenaustausch |q application/pdf |u http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027180093&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
| 912 | |a ebook | ||
| 999 | |a oai:aleph.bib-bvb.de:BVB01-027180093 | ||
Datensatz im Suchindex
| _version_ | 1804152017828970496 |
|---|---|
| adam_text | CONTENTS II
1 INTRODUCTION 1
1.1 BACKGROUND AND AIM OF THE STUDY 2
1.2 NATURAL COMPOUNDS 4
1.2.1 IMPORTANCE OF NATURAL COMPOUNDS 4
1.2.2 MYXOBACTERIA AS PRODUCERS OF NATURAL COMPOUNDS 4
1.2.3 THE MYXOBACTERIAL COMPOUND PRETUBULYSIN 5
1.3 THE MICROTUBULE SYSTEM 5
1.3.1 STRUCTURE OF MICROTUBULES 6
1.3.2 DYNAMIC MICROTUBULES
- MODE OF ACTION 7
1.3.3 REGULATION OF MICROTUBULE DYNAMICS 8
1.3.4 NATURAL PRODUCTS THAT TARGET
MICROTUBULES 8
1.3.5 THE BIG PROBLEM - RESISTANCE TO MTAS 9
1.4 ANTI-VASCULAR STRATEGIES FOR TUMOR
TREATMENT 10
1.4.1 NORMAL VASCULATURE VS. TUMOR VASCULATURE 10
1.4.2 ANGIOGENESIS INHIBITORS AND VASCULAR DISRUPTING AGENTS 12
1.4.3 CLASSES OF VDAS 12
1.4.4 MODE OF ACTION OF MICROTUBULE TARGETING VDAS 13
1.4.4.1 EFFECTS ON TUMOR VASCULATURE 13
1.4.4.2 EFFECTS ON TUMOR EC 14
1.4.5 LIMITATIONS OF VDAS 16
2 MATERIALS AND METHODS 17
2.1 MATERIALS 18
2.1.1 COMPOUNDS 18
2.1.2 BIOCHEMICALS, DYES, INHIBITORS AND CELL CULTURE REAGENTS 18
2.2 CELL CULTURE 20
2.2.1 BUFFERS, SOLUTIONS AND REAGENTS 20
2.2.2 CELL LINES 22
2.2.2.1 HUVECS - HUMAN UMBILICAL VEIN ENDOTHELIAL CELLS 22
2.2.2.2 HMEC-1
- HUMAN DERMAL
MICROVASCULAR ENDOTHELIAL CELLS 22
2.2.2.3 HAMSTER A-MEL-3 AMELANOTIC MELANOMA CELL LINE 23
2.2.2.4 B16-F1
- MOUSE SKIN MELANOMA CELL LINE 23
HTTP://D-NB.INFO/104778047X
CONTENTS III
2.2.3 PASSAGING 23
2.2.4 FREEZING AND THAWING 24
2.3 WESTERN BLOT ANALYSIS 24
2.3.1 SAMPLE PREPARATION 24
2.3.2 PROTEIN QUANTIFICATION 25
2.3.2.1 BRADFORD ASSAY 25
2.3.2.2 BICINCHONINIC (BCA) PROTEIN ASSAY 25
2.3.3 SDS-PAGE 26
2.3.4 TANK ELECTROBLOTTING 27
2.3.5 PROTEIN DETECTION 27
2.3.5.1 CONTROL-STAINING OF POLYACRYLAMIDE GELS 27
2.3.5.2 PROTEIN DETECTION
ON NITROCELLULOSE MEMBRANES 28
2.3.5.2.1 ENHANCED CHEMILUMINESCENCE 28
2.3.5.2.2 INFRARED IMAGING 28
2.4 ACTIVE RHOA PULL-DOWN ASSAY 30
2.5 IMMUNOCYTOCHEMISTRY AND CONFOCAL LASER SCANNING MICROSCOPY 30
2.5.1 F-ACTIN, PP-MLC2, AND
EC JUNCTION STAINING 30
2.5.2 MICROTUBULE STAINING 31
2.6 PERMEABILITY ASSAYS 32
2.6.1 IMPEDANCE MEASUREMENT 32
2.6.2 MACROMOLECULAR PERMEABILITY ASSAY 33
2.7 CELL VIABILITY ASSAYS 33
2.7.1 CELLTITER-BLUE CELL VIABILITY ASSAY 33
2.7.2 FLOW CYTOMETRY 34
2.7.2.1 QUANTIFICATION OF APOPTOTIC CELL DEATH 34
2.7.2.2 CELL CYCLE ANALYSIS 35
2.7.2.3 QUANTIFICATION OF NECROTIC CELL DEATH 35
2.7.3 MONITORING OF MORPHOLOGICAL CHANGES 36
2.8 VE-CADHERIN QUANTIFICATION VIA
FACS ANALYSIS 36
2.9 ENDOTHELIAL TUBE DISRUPTION ASSAYS 36
2.9.1 TUBE DISRUPTION
(IN VITRO) 36
2.9.2 MOUSE AORTIC RING ASSAY (EX VIVO) 37
CONTENTS IV
2.10 CYTOSOLIC CALCIUM IMAGING 37
2.11 IN VIVO EXPERIMENTS 38
2.11.1 ANIMALS 38
2.11.2 DETERMINING THE MAXIMUM TOLERATED DOSE (MTD) OF PT 38
2.11.3 HAMSTER DORSAL SKINFOLD CHAMBER MEASUREMENT 39
2.11.4 B16-F1 MOUSE MELANOMA TUMOR MODEL (SINGLE DOSE) 40
2.11.4.1 QUANTIFICATION OF HOECHST 33342 PERFUSION AND VESSEL DENSITY 40
2.11.4.2 HAEMATOXYLIN AND EOSIN STAINING (H&E) 41
2.11.5 B16-F1 MOUSE MELANOMA TUMOR MODEL (MULTIPLE DOSES) 41
2.12 STATISTICAL ANALYSIS 41
3 RESULTS 42
3.1 PT INDUCES TYPICAL
HALLMARKS OF VASCULAR DISRUPTION IN ECS IN VITRO 43
3.1.1 PT INDUCES THE DEPOLYMERIZATION OF MICROTUBULES 43
3.1.2 PT TRIGGERS ACTIN STRESS FIBER FORMATION, DISRUPTS EC JUNCTIONS,
AND INDUCES
REORGANIZATION OF FOCAL ADHESIONS 44
3.1.3 PT RAPIDLY INDUCES ENDOTHELIAL BARRIER BREAKDOWN 45
3.1.3.1 IMPEDANCE SENSING 45
3.1.3.2 MACROMOLECULAR PERMEABILITY 46
3.1.3.3 PT REDUCES EXTRACELLULAR VE-CADHERIN EXPRESSION 46
3.1.4 PT DISRUPTS ESTABLISHED ENDOTHELIAL TUBES 47
3.1.4.1 PT DISRUPTS ENDOTHELIAL TUBES IN VITRO 47
3.1.4.2 PT DISRUPTS ENDOTHELIAL SPROUTS EX VIVO 49
3.1.5 PT TREATMENT IS NOT CYTOTOXIC FOR ECS 50
3.1.5.1 METABOLIC ACTIVITY 50
3.1.5.2 QUANTIFICATION OF APOPTOTIC AND NECROTIC CELLS 50
3.1.5.3 RECOVERY AFTER WITHDRAWAL OF PT 52
3.2 PT-INDUCED SIGNALING CASCADES LEADING TO ACTIN STRESS
FIBER FORMATION
AND HYPERPERMEABILITY 53
3.2.1 INVOLVEMENT OF THE RHOA/ROCK/MLC PATHWAY 54
3.2.1.1 RHOA 54
3.2.1.2 MLC2 55
3.2.1.3 MLCP AND MLCK 55
CONTENTS V
3.2.1.4 ROCK 56
3.2.2 INVOLVEMENT OF MAPK PATHWAYS 58
3.2.3 P38 MAPK AND JNK 58
3.2.4 INVOLVEMENT OF INTRACELLULAR CALCIUM
[CA
2+
]J
60
3.3 SINGLE DOSE TREATMENT WITH PT SELECTIVELY AFFECTS TUMOR VASCULATURE
IN VIVO 62
3.3.1 DETERMINING THE MAXIMUM TOLERATED DOSE (MTD) OF PT 62
3.3.2 PT SELECTIVELY DIMINISHED TUMOR BLOOD FLOW IN A HAMSTER DORSAL
SKINFOLD
CHAMBER MODEL 63
3.3.3 PT REDUCES TUMOR VESSEL PERFUSION WITHOUT AFFECTING THE DENSITY OF
TUMOR
VESSELS IN AN ECTOPIC B16-F1 MOUSE MELANOMA TUMOR MODEL 64
3.3.4 PT INDUCES ENORMOUS CENTRAL NECROSIS IN B16-F1 MOUSE MELANOMA
TUMORS 66
3.4 MULTIPLE DOSE TREATMENT OF PT DECELERATES TUMOR GROWTH IN VIVO 67
4 DISCUSSION 68
4.1 PT TREATMENT ELICITS TYPICAL HALLMARKS OF VASCULAR DISRUPTION IN ECS
IN VITRO 69
4.1.1 THE IMPACT OF PT ON MICROTUBULES 69
4.1.2 THE LINK BETWEEN PT-MEDIATED MICROTUBULE DEPOLYMERIZATION AND
ACTIN
STRESS FIBER FORMATION 70
4.1.3 PT AND THE MICROTUBULE MEDIATED PROCESS OF MEMBRANE BLEBBING 71
4.1.4 THE INFLUENCE OF PT ON ENDOTHELIAL JUNCTIONS 72
4.1.5 CYTOTOXIC PROFILE OF PT TREATMENT IN CONFLUENT ECS 73
%
4.2 PT SELECTIVELY TARGETS TUMOR VASCULATURE IN VIVO 74
4.2.1 MECHANISMS LEADING TO SELECTIVE BLOOD FLOW SHUTDOWN 74
4.2.2 THERAPEUTIC POTENTIAL OF PT 75
4.2.3 POSSIBLE ADVERSE EFFECTS OF PT TREATMENT 75
5 SUMMARY AND CONCLUSION 77
6 REFERENCES 79
7 APPENDIX 91
CONTENTS V[
7.1 PUBLICATIONS 92
7.1.1 ORIGINAL PUBLICATIONS 92
7.1.2 ORAL PRESENTATIONS 92
7.1.3 POSTER PRESENTATIONS 93
7.2 CURRICULUM VITAE 94
7.3 ACKNOWLEDGEMENTS 95
|
| any_adam_object | 1 |
| author | Kretzschmann, Verena |
| author_facet | Kretzschmann, Verena |
| author_role | aut |
| author_sort | Kretzschmann, Verena |
| author_variant | v k vk |
| building | Verbundindex |
| bvnumber | BV041733356 |
| collection | ebook |
| ctrlnum | (OCoLC)876897736 (DE-599)DNB1046503227 |
| discipline | Medizin |
| format | Thesis Book |
| fullrecord | <?xml version="1.0" encoding="UTF-8"?><collection xmlns="http://www.loc.gov/MARC21/slim"><record><leader>01789nam a2200409zc 4500</leader><controlfield tag="001">BV041733356</controlfield><controlfield tag="003">DE-604</controlfield><controlfield tag="005">20140327 </controlfield><controlfield tag="007">t</controlfield><controlfield tag="008">140313s2013 gw ad|| m||| 00||| eng d</controlfield><datafield tag="015" ind1=" " ind2=" "><subfield code="a">14,O02</subfield><subfield code="2">dnb</subfield></datafield><datafield tag="016" ind1="7" ind2=" "><subfield code="a">1046503227</subfield><subfield code="2">DE-101</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(OCoLC)876897736</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(DE-599)DNB1046503227</subfield></datafield><datafield tag="040" ind1=" " ind2=" "><subfield code="a">DE-604</subfield><subfield code="b">ger</subfield><subfield code="e">rakddb</subfield></datafield><datafield tag="041" ind1="0" ind2=" "><subfield code="a">eng</subfield></datafield><datafield tag="044" ind1=" " ind2=" "><subfield code="a">gw</subfield><subfield code="c">XA-DE-BY</subfield></datafield><datafield tag="049" ind1=" " ind2=" "><subfield code="a">DE-12</subfield><subfield code="a">DE-19</subfield><subfield code="a">DE-384</subfield><subfield code="a">DE-473</subfield><subfield code="a">DE-703</subfield><subfield code="a">DE-1051</subfield><subfield code="a">DE-824</subfield><subfield code="a">DE-29</subfield><subfield code="a">DE-91</subfield><subfield code="a">DE-1049</subfield><subfield code="a">DE-92</subfield><subfield code="a">DE-739</subfield><subfield code="a">DE-898</subfield><subfield code="a">DE-355</subfield><subfield code="a">DE-706</subfield><subfield code="a">DE-20</subfield><subfield code="a">DE-1102</subfield></datafield><datafield tag="084" ind1=" " ind2=" "><subfield code="a">610</subfield><subfield code="2">sdnb</subfield></datafield><datafield tag="100" ind1="1" ind2=" "><subfield code="a">Kretzschmann, Verena</subfield><subfield code="e">Verfasser</subfield><subfield code="4">aut</subfield></datafield><datafield tag="245" ind1="1" ind2="0"><subfield code="a">Preclinical characterization of the myxobacterial compound pretubulysin as novel vascular disrupting agent</subfield><subfield code="c">Verena Kretzschmann</subfield></datafield><datafield tag="264" ind1=" " ind2="1"><subfield code="c">2013</subfield></datafield><datafield tag="300" ind1=" " ind2=" "><subfield code="a">VI, 96 Blatt</subfield><subfield code="b">Ill., graph. Darst.</subfield></datafield><datafield tag="336" ind1=" " ind2=" "><subfield code="b">txt</subfield><subfield code="2">rdacontent</subfield></datafield><datafield tag="337" ind1=" " ind2=" "><subfield code="b">n</subfield><subfield code="2">rdamedia</subfield></datafield><datafield tag="338" ind1=" " ind2=" "><subfield code="b">nc</subfield><subfield code="2">rdacarrier</subfield></datafield><datafield tag="502" ind1=" " ind2=" "><subfield code="a">München, Univ., Diss., 2013</subfield></datafield><datafield tag="538" ind1=" " ind2=" "><subfield code="a">Langzeitarchivierung gewährleistet, LZA</subfield></datafield><datafield tag="655" ind1=" " ind2="7"><subfield code="0">(DE-588)4113937-9</subfield><subfield code="a">Hochschulschrift</subfield><subfield code="2">gnd-content</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Vollmar, Angelika</subfield><subfield code="4">dgs</subfield></datafield><datafield tag="776" ind1="0" ind2="8"><subfield code="i">Erscheint auch als</subfield><subfield code="n">Online-Ausgabe</subfield><subfield code="o">urn:nbn:de:bvb:19-164317</subfield></datafield><datafield tag="856" ind1="4" ind2="0"><subfield code="u">https://nbn-resolving.org/urn:nbn:de:bvb:19-164317</subfield><subfield code="x">Resolving-System</subfield><subfield code="z">kostenfrei</subfield><subfield code="3">Volltext</subfield></datafield><datafield tag="856" ind1="4" ind2="0"><subfield code="u">http://d-nb.info/1046503227/34</subfield><subfield code="x">Langzeitarchivierung Nationalbibliothek</subfield></datafield><datafield tag="856" ind1="4" ind2="0"><subfield code="u">http://edoc.ub.uni-muenchen.de/16431/</subfield><subfield code="x">Verlag</subfield><subfield code="z">kostenfrei</subfield></datafield><datafield tag="856" ind1="4" ind2="2"><subfield code="m">DNB Datenaustausch</subfield><subfield code="q">application/pdf</subfield><subfield code="u">http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027180093&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA</subfield><subfield code="3">Inhaltsverzeichnis</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">ebook</subfield></datafield><datafield tag="999" ind1=" " ind2=" "><subfield code="a">oai:aleph.bib-bvb.de:BVB01-027180093</subfield></datafield></record></collection> |
| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV041733356 |
| illustrated | Illustrated |
| indexdate | 2024-07-10T01:04:02Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-027180093 |
| oclc_num | 876897736 |
| open_access_boolean | 1 |
| owner | DE-12 DE-19 DE-BY-UBM DE-384 DE-473 DE-BY-UBG DE-703 DE-1051 DE-824 DE-29 DE-91 DE-BY-TUM DE-1049 DE-92 DE-739 DE-898 DE-BY-UBR DE-355 DE-BY-UBR DE-706 DE-20 DE-1102 |
| owner_facet | DE-12 DE-19 DE-BY-UBM DE-384 DE-473 DE-BY-UBG DE-703 DE-1051 DE-824 DE-29 DE-91 DE-BY-TUM DE-1049 DE-92 DE-739 DE-898 DE-BY-UBR DE-355 DE-BY-UBR DE-706 DE-20 DE-1102 |
| physical | VI, 96 Blatt Ill., graph. Darst. |
| psigel | ebook |
| publishDate | 2013 |
| publishDateSearch | 2013 |
| publishDateSort | 2013 |
| record_format | marc |
| spelling | Kretzschmann, Verena Verfasser aut Preclinical characterization of the myxobacterial compound pretubulysin as novel vascular disrupting agent Verena Kretzschmann 2013 VI, 96 Blatt Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier München, Univ., Diss., 2013 Langzeitarchivierung gewährleistet, LZA (DE-588)4113937-9 Hochschulschrift gnd-content Vollmar, Angelika dgs Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:19-164317 https://nbn-resolving.org/urn:nbn:de:bvb:19-164317 Resolving-System kostenfrei Volltext http://d-nb.info/1046503227/34 Langzeitarchivierung Nationalbibliothek http://edoc.ub.uni-muenchen.de/16431/ Verlag kostenfrei DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027180093&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Kretzschmann, Verena Preclinical characterization of the myxobacterial compound pretubulysin as novel vascular disrupting agent |
| subject_GND | (DE-588)4113937-9 |
| title | Preclinical characterization of the myxobacterial compound pretubulysin as novel vascular disrupting agent |
| title_auth | Preclinical characterization of the myxobacterial compound pretubulysin as novel vascular disrupting agent |
| title_exact_search | Preclinical characterization of the myxobacterial compound pretubulysin as novel vascular disrupting agent |
| title_full | Preclinical characterization of the myxobacterial compound pretubulysin as novel vascular disrupting agent Verena Kretzschmann |
| title_fullStr | Preclinical characterization of the myxobacterial compound pretubulysin as novel vascular disrupting agent Verena Kretzschmann |
| title_full_unstemmed | Preclinical characterization of the myxobacterial compound pretubulysin as novel vascular disrupting agent Verena Kretzschmann |
| title_short | Preclinical characterization of the myxobacterial compound pretubulysin as novel vascular disrupting agent |
| title_sort | preclinical characterization of the myxobacterial compound pretubulysin as novel vascular disrupting agent |
| topic_facet | Hochschulschrift |
| url | https://nbn-resolving.org/urn:nbn:de:bvb:19-164317 http://d-nb.info/1046503227/34 http://edoc.ub.uni-muenchen.de/16431/ http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027180093&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT kretzschmannverena preclinicalcharacterizationofthemyxobacterialcompoundpretubulysinasnovelvasculardisruptingagent AT vollmarangelika preclinicalcharacterizationofthemyxobacterialcompoundpretubulysinasnovelvasculardisruptingagent |