SRC a critical signaling mediator in FLT 3 ITD positive AML:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2012
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| Schlagworte: | |
| Online-Zugang: | Volltext https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20120321-1118166-0-6 Inhaltsverzeichnis |
| Beschreibung: | X, 105 Bl. Ill., graph. Darst. |
Internformat
MARC
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| 100 | 1 | |a Leischner, Hannes |d 1982- |e Verfasser |0 (DE-588)135617596 |4 aut | |
| 245 | 1 | 0 | |a SRC a critical signaling mediator in FLT 3 ITD positive AML |c Hannes Leischner |
| 264 | 1 | |c 2012 | |
| 300 | |a X, 105 Bl. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
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| 502 | |a München, Techn. Univ., Diss., 2012 | ||
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| 856 | 4 | 1 | |u http://mediatum.ub.tum.de/node?id=1118166 |x Verlag |z kostenfrei |3 Volltext |
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| 912 | |a ebook | ||
| 999 | |a oai:aleph.bib-bvb.de:BVB01-025876077 | ||
Datensatz im Suchindex
| _version_ | 1804150180635738112 |
|---|---|
| adam_text | IMAGE 1
CONTENT
CONTENT I
ABBREVIATIONS VII
1 SUMMARY 1
2 INTRODUCTION 3
2.1 THE ACUTE MYELOID LEUKEMIA (AML) 3
2.1.2 AML THERAPY 5
2.2 THE PROTEIN FLT3 6
2.2.1 STRUCTURE AND FUNCTION OF FMS-LIKE TYROSINE KINASE 3 6
2.2.2 FLT3 FUNCTION IN NORMAL AND MALIGNANT HEMATOPOIESIS 7
2.2.3 EXPRESSION IN HEMATOLOGIC MALIGNANCIES 8
2.2.4 ALTERATIONS OF FLT3 IN AML 9
2.2.5 FLT3 INTERNAL TANDEM DUPLICATION MUTATIONS (FLT3 ITD) 9
2.2.6 FLT3 ACTIVATION LOOP MUTATION (FLT3 TKD) 10
2.2.7 CLINICAL SIGNIFICANCE OF FLT3 ITD 11
2.2.8 SIGNALING OF FLT3 AND ACTIVATED PATHWAYS 11
2.2.9 FLT3 BASED AML TREATMENT APPROACHES 13
2.3 THE SRC FAMILY KINASE FAMILY (SFK) 14
2.3.1 SRC STRUCTURE AND FUNCTION 14
2.3.2 INVOLVEMENT OF SRC IN HUMAN CANCERS 16
2.3.3 SFKS IN LEUKEMIAS, LYMPHOMAS, AND MYELOMAS 16
2.3.4 SRC KINASE INHIBITORS 17
3 AIM OF THE STUDY 20
4 RESULTS 21
I
HTTP://D-NB.INFO/1035257866
IMAGE 2
CONTENT
4.1 CHARACTERIZATION OF FLT3 ITD AND FLT3 TKD SIGNALING 21
4.2 TESTING OF SFK AS POTENTIAL FLT3 ITD INTERACTION PARTNERS 23
4.3 INTERACTION OF SRC AND FLT3 WT, FLT3 ITD AND FLT3 TKD 24
4.4 INTERACTION OF SRC AND FLT3 ITD 26
4.5 IDENTIFICATION OF Y589 AND Y591 AS BINDING SITES OF SRC AND FLT3 ITD
26
4.6 Y589 AND Y591 SHOW DIFFERENT PATTERNS OF PHOSPHORYLATION IN FLT3 WT,
FLT3
ITD OR FLT3TKD 28
4.7 THE INHIBITOR PD166-326 HAS GROWTH INHIBITORY EFFECTS ON FLT3 ITD
CELLS ....29
4.8 PD166-326 LEADS TO REDUCED PSTAT5 LEVELS IN FLT3 ITD CELLS THROUGH
SRC
INHIBITION 30
4.9 COMBINATION PD166-326 AND PKC412 HAS ADDITIVE INHIBITORY EFFECTS ON
FLT3
ITD POSITIVE CELLS 32
4.10 THE BIOCHEMICAL EFFECT OF PKC412 AND PD166-326 COMBINATION ON
DOWNSTREAM TARGETS OF FLT3 ITD 34
4.11 SIRNA MEDIATED DOWN-REGULATION OF SRC LEADS TO REDUCED
PROLIFERATION OF
FLT3 ITD POSITIVE CELLS 34
4.12 ASSESSMENT OF THE BIOCHEMICAL EFFECT OF SRC DOWN-REGULATION ON FLT3
ITD
AND FLT3 TKD POSITIVE CELLS 36
4.13 THE EFFECT OF REDUCED SRC EXPRESSION ON FLT3 ITD INDUCED LEUKEMIA
IN
MICE 37
4.14 LCK IS NOT INVOLVED IN THE SIGNAL TRANSDUCTION OF FLT3 ITD 38
4.15 COMBINATION OF PKC412 AND DASATINIB HAS INHIBITORY EFFECTS ON HUMAN
AND
MURINE FLT3 ITD POSITIVE CELLS 40
4.16 BIOCHEMICAL EFFECT OF THE COMBINATION OF PKC412 AND DASATINIB ON
MV4-11
CELLS 41
4.17 COMBINATION OF PKC412 AND DASATINIB INCREASES APOPTOSIS IN MV4-11
CELLS.
42
4.18 DASATINIB AND PKC412 SHOW ADDITIVE INHIBITORY EFFECTS ON PRIMARY
HUMAN
AML CELLS 43
5 DISCUSSION 45
5.1 IDENTIFICATION OF POTENTIAL FLT3 ITD INTERACTING PARTNERS 45
5.2 THE ROLE OF Y589 AND Y591 FOR THE INTERACTION OF FLT3 ITD AND SRC 47
II
IMAGE 3
CONTENT
5.3 THE PHOSPHORYLATION OF Y589 AND Y591 DIFFERS BETWEEN FLT3 WT AND
MUTANTS 48
5.4 THE EFFECT OF REDUCED SRC EXPRESSION ON FLT3 ITD AND FLT3 TKD
POSITIVE
CELLS 49
5.5 THE INFLUENCE ON SRC DOWN REGULATION ON FLT3 ITD INDUCED LEUKEMIA IN
MICE 49
5.6 FLT3 MUTANTS RESPOND DIFFERENTLY TO SRC INHIBITORS AND INHIBITOR
COMBINATIONS 50
5.7 SRC AS A THERAPEUTIC TARGET IN HUMAN AML THERAPY 51
6 MATERIAL 53
6.1 REAGENTS 53
6.2 MEDIUM AND SUPPLEMENTS FOR CELL CULTURE 55
6.3 ENZYMES 55
6.4 MEMBRANES 56
6.5 ANTIBODIES 56
6.6 MATERIAL AND KITS 57
6.7 DNA AND PROTEIN MARKER 57
6.8 INSTRUMENTS 58
6.9 OLIGONUCLEOTIDES 59
6.9.1 SHRNA SEQUENCES 59
6.10 DNA-CONSTRUCTS 60
6.11 BACTERIAS 61
6.12 CELL LINES 61
6.13 MICE 61
6.14 MATERIALS USED IN THE ANIMAL EXPERIMENTS 62
6.15 SOLUTIONS AND BUFFER 62
7 METHODS 67
7.1 MOLECULAR BIOLOGY TECHNIQUES 67
7.1.1 POLYMERASE CHAIN REACTION 67
7.1.2 DIGESTION OF DNA WITH RESTRICTION ENZYMES 67
7.1.3 AGAROSE GEL ELECTROPHORESIS 68
III
IMAGE 4
CONTENT
7.1.4 DEPHOSPHORYLATION OF 5 -PHOSPHATE RESIDUES OF DNA USING CALF
INTESTINE
PHOSPHATASE 68
7.1.5 GEL EXTRACTION 69
7.1.6 LIGATION 69
7.2 WORKING WITH BACTERIA 69
7.2.1 PREPARATION OF THE COMPETENT BACTERIA 69
7.2.2 TRANSFORMATION OF COMPETENT BACTERIA 70
7.2.3 PLASMID-PREPARATION 70
7.2.4 SEQUENCING 71
7.3 CELL CULTURE 71
7.3.1 CULTURING OF SUSPENSION CELLS 71
7.3.2 CULTURING OF ADHERENT CELLS 72
7.3.3 FREEZING OF CELLS 72
7.3.4 THAWING OF CELLS 72
7.3.5 CELL QUANTIFICATION AND EVALUATION OF VIABILITY 73
7.3.6 STARVING OF THE CELLS PRIOR LYSIS 73
7.3.7 STIMULATION OF CELLS WITH CYTOKINE/LIGAND BEFORE CELL LYSIS 73
7.3.8 TRANSFECTION 73
7.3.9 VIRUS PRODUCTION AND RETROVIRAL TRANSDUCTION 74
7.3.10 FACS-ANALYSIS 76
7.3.11 SELECTION OF TRANSDUCED CELLS 76
7.3.12 PREPARATION OF CELL LYSATES 76
7.3.13 GST PURIFICATION AND GST BINDING STUDIES 77
7.3.14 IMMUNOPRECIPITATION OF PROTEINS FROM CELLULAR LYSATES 77
7.3.15 SDS-PAGE ELECTROPHORESIS 78
7.3.16 WESTERN BLOT 78
7.3.17 IMMUNHISTOCHEMICAL DETECTION OF PROTEINS 79
7.4 BIOLOGICAL ASSAYS 79
7.4.1 APOPTOSIS ASSAY USING ANNEXIN-V APOPTOSIS DETECTION KIT 79
7.4.2 PROLIFERATION ASSAY 80
7.5 ANIMAL EXPERIMENTS 80
7.5.1 ANIMAL CARE 80
IV
IMAGE 5
CONTENT
7.5.2 PREPARATION OF THE CELLS AND MICE PRIOR TO TRANSPLANTATION 81
7.5.3 DRAWING OF BLOOD 81
7.5.4 MONITORING OF THE TRANSPLANTED MICE 81
8 LITERATURE 83
9 FIGURES 100
10 ACKNOWLEDGEMENT 103
11 PUBLICATIONS 105
V
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| physical | X, 105 Bl. Ill., graph. Darst. |
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| spelling | Leischner, Hannes 1982- Verfasser (DE-588)135617596 aut SRC a critical signaling mediator in FLT 3 ITD positive AML Hannes Leischner 2012 X, 105 Bl. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier München, Techn. Univ., Diss., 2012 (DE-588)4113937-9 Hochschulschrift gnd-content Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:91-diss-20120321-1118166-0-6 http://mediatum.ub.tum.de/node?id=1118166 Verlag kostenfrei Volltext https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20120321-1118166-0-6 Resolving-System DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025876077&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Leischner, Hannes 1982- SRC a critical signaling mediator in FLT 3 ITD positive AML |
| subject_GND | (DE-588)4113937-9 |
| title | SRC a critical signaling mediator in FLT 3 ITD positive AML |
| title_auth | SRC a critical signaling mediator in FLT 3 ITD positive AML |
| title_exact_search | SRC a critical signaling mediator in FLT 3 ITD positive AML |
| title_full | SRC a critical signaling mediator in FLT 3 ITD positive AML Hannes Leischner |
| title_fullStr | SRC a critical signaling mediator in FLT 3 ITD positive AML Hannes Leischner |
| title_full_unstemmed | SRC a critical signaling mediator in FLT 3 ITD positive AML Hannes Leischner |
| title_short | SRC a critical signaling mediator in FLT 3 ITD positive AML |
| title_sort | src a critical signaling mediator in flt 3 itd positive aml |
| topic_facet | Hochschulschrift |
| url | http://mediatum.ub.tum.de/node?id=1118166 https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20120321-1118166-0-6 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025876077&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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