Innovative therapeutic approaches in chronic lymphocytic leukemia:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
Freiburg im Breisgau
Oktober 2021
|
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| Beschreibung: | 171 Seiten Illustrationen, Diagramme 30 cm |
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| 100 | 1 | |a Khaja Saleem, Al Ahmirathus Syedush Shifa |d 1990- |e Verfasser |0 (DE-588)126068105X |4 aut | |
| 245 | 1 | 0 | |a Innovative therapeutic approaches in chronic lymphocytic leukemia |c vorgelegt von Al Ahmirathus Syedush Shifa Khaja Saleem |
| 264 | 1 | |a Freiburg im Breisgau |c Oktober 2021 | |
| 300 | |a 171 Seiten |b Illustrationen, Diagramme |c 30 cm | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |b Dissertation |c Albert-Ludwigs-Universität Freiburg im Breisgau |d 2022 | ||
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 856 | 4 | 2 | |m B:DE-101 |q application/pdf |u https://d-nb.info/1264509839/04 |3 Inhaltsverzeichnis |
| 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=034565569&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
| 999 | |a oai:aleph.bib-bvb.de:BVB01-034565569 | ||
| 883 | 0 | |8 1\p |a emakn |c 0,83402 |d 20220812 |q DE-101 |u https://d-nb.info/provenance/plan#emakn | |
| 883 | 2 | |8 2\p |a dnb |d 20220909 |q DE-101 |u https://d-nb.info/provenance/plan#dnb | |
Datensatz im Suchindex
| _version_ | 1804185803440521216 |
|---|---|
| adam_text | TABLE
OF
CONTENTS
1
SUMMARY
....................................................................................................
5
1.1
SUMMARY
I
..........................................................................................................
5
1.2
SUMMARY
II
.........................................................................................................
6
2
INTRODUCTION
...............................................................................................
7
2.1
CHRONIC
LYMPHOCYTIC
LEUKEMIA
.........................................................................
7
2.1.1
CHARACTERISTICS
OF
CLL
.................................................................................
7
2.1.2
CLINICAL
STAGING
AND
PROGNOSTIC
MARKERS
IN
CLL
..........................................
7
2.2
CLL
MICROENVIRONMENT
.......................................................................................
9
2.3
CELL
SIGNALING
IN
CLL
.........................................................................................
12
2.3.1
B-CELL
RECEPTOR
PATHWAY
............................................................................
12
2.3.2
JNK1
SIGNALING
..........................................................................................
22
2.3.3
NOTCH
AND
NFKB
SIGNALING
......................................................................
23
2.4
THERAPEUTIC
OPTIONS
IN
B-CLL
CELL
RELATED
IMMUNOLOGICAL
DISEASES
.................
24
2.4.1
SIGNALING
TARGETS
FOR
CLL
THERAPY
..............................................................
24
2.4.2 EPIGENETIC
TARGETED
THERAPIES
IN
CLL
..........................................................
31
3
AIMS
OF
THE
STUDY
....................................................................................
34
4
MATERIALS
...................................................................................................
35
4.1
LABORATORY
EQUIPMENTS
AND
DISPOSABLES
..........................................................
35
4.1.1
EQUIPMENT
..................................................................................................
35
4.1.2
DISPOSABLES
...............................................................................................
37
4.1.3
CHEMICALS
AND
REAGENTS
............................................................................
38
4.1.4
BUFFERS
AND
SOLUTIONS
.................................................................................
41
4.2
MEDIA
................................................................................................................
43
4.3
KITS
...................................................................................................................
44
4.4
HUMAN
BLOOD
SAMPLES
.....................................................................................
44
4.4.1.
PRIMARY
HUMAN
CLL
SAMPLES
...................................................................
44
1
4.4.2.
HEALTHY
DONOR
BLOOD
SAMPLES
...................................................................
44
4.5
CELL
LINES
...........................................................................................................
45
4.6
ANTIBODIES
.........................................................................................................
45
4.6.1.
FACS
........................................................................................................
45
4.6.2.
CO-IMMUNOPRECIPITATION
AND
WESTERNBLOT
..................................................
46
4.7
.
INHIBITORS
AND
SIRNA
.................................................................................
50
4.7.1.
JNK1
AND
BROMODOMAIN
INHIBITORS
...........................................................
50
4.7.2.
SIRNAS
.....................................................................................................
50
4.8
.
OLIGONUCLEOTIDES
........................................................................................
51
4.9
.
MOUSE
STRAINS
AND
ANIMAL
HUSBANDRY
........................................................
51
4.10
.
DATABASES
...............................................................................................
52
4.11
.
SOFTWARE
AND
TOOLS
...................................................................................
52
5
METHODS
....................................................................................................
54
5.1.
IN
VITRO
EXPERIMENTS
..........................................................................................
54
5.1.1.
CELL
CULTURE
................................................................................................
54
5.1.2.
B
CELL
ISOLATION
...........................................................................................
55
5.1.3.
INHIBITORS
TREATMENT
....................................................................................
55
5.1.4.
SIRNA
MEDIATED
KNOCK
DOWN
....................................................................
56
5.1.5.
APOPTOSIS
ASSAY
........................................................................................
56
5.1.6.
PROLIFERATION
ASSAY
.....................................................................................
57
5.1.7.
TRANSWELL
MIGRATION
ASSAY
........................................................................
58
5.1.8.
CXCR4
SURFACE
EXPRESSION
......................................................................
58
5.1.9.
PROTEIN
ANALYSIS
.........................................................................................
58
5.1.10.
CELL
CYCLE
ANALYSIS
..................................................................................
60
5.1.11.
RNA
EXTRACTION,
REVERSE
TRANSCRIPTION
AND
QPCR
(TAQMAN)
.................
60
5.1.12.
GENE
ARRAY
(BY
DR.
DIETMAR
PFEIFER)
........................................................
61
5.2.
IN
VIVO
EXPERIMENTS
..........................................................................................
61
5.2.1
XENOGRAFT
MOUSE
MODEL
............................................................................
61
5.2.2.
EP-TCL1
MOUSE
MODEL
............................................................................
61
2
5.2.3.
TREATMENT
EXPERIMENTS
.............................................................................
62
5.2.4.
BLOOD
WITHDRAWAL
AND
PROCESSING
OF
BLOOD
SAMPLES
.................................
63
5.2.5.
MOUSE
FOLLOW-UP
AND
SACRIFICE
THE
MICE
AFTER
DRUG
THERAPY
......................
63
5.2.6.
FACS
........................................................................................................
64
5.2.7.
IMMUNOHISTOCHEMISTRY
(BY
DR.
KONRAD
AUMANN)
.....................................
73
5.3.
IN
SILICO
ANALYSIS
(BY
DR.
DIETMAR
PFEIFER)
.........................................................
73
5.4.
STATISTICAL
ANALYSIS
...........................................................................................
73
6
RESULTS
.....................................................................................................
74
6.1
RESULTS
1
............................................................................................................
74
6.1.1.
JNK1
IS
OVEREXPRESSED
AND
HYPERPHOSPHORYLATED
IN
HUMAN
PRIMARY
CLL
CELLS
74
6.1.2.
JNK1
KNOCK-DOWN
AND
JNK1
INHIBITION
AFFECTS
PRIMARY
CLL
CELL
VIABILITY.
76
6.1.3.
JNK1
INHIBITORS
REDUCE
ANTI-APOPTOTIC
DOWNSTREAM
EFFECTORS
IN
CLL
......
80
6.1.4.
SP600125
INHIBITOR
INDUCES
APOPTOSIS
AND
AFFECTS
THE
DOWNSTREAM
TARGETS
IN
UNMUTATED
CLL
IN
THE
PRESENCE
OF
PROTECTIVE
STROMAL
CELLS
............................
81
6.1.5.
JNK1
IS
A
CRUCIAL
DOWNSTREAM
MEDIATOR
OF
B-CELL
RECEPTOR
SIGNALING
IN
CLL
82
6.1.6.
JNK1
INHIBITION
REDUCES
LEUKEMIC
BURDEN
IN
SPLEEN
AND
BLOOD
OF
CLL
TRANSPLANTED
NOG
MICE
.....................................................................................
85
6.1.7.
JNK1
INHIBITOR
DECREASES
LEUKEMIC
CLL
CELLS
AND
MODULATES
IMMUNE
RESPONSES
IN
EP-TCL1
MOUSE
MODEL
................................................................
89
6.2
RESULTS
II
...........................................................................................................
94
6.2.1
FUNCTIONAL
ANALYSIS
AFTER
BET
INHIBITION
IN
B-CLL
CELL
LINE
AND
PRIMARY
CLL
CELLS
94
6.2.2
BET
INHIBITOR
AFFECTS
STAT5
AND
OTHER
PROLIFERATIVE
PROTEINS
INVOLVE
IN
CLL
CELL
PROLIFERATION
AND
SURVIVAL
...............................................................................
98
6.2.3
JQ1
AFFECTS
MIGRATORY
EFFECT
OF
CLL
CELLS
THROUGH
REGULATION
OF
STAT5
DEPENDENT
AND
INDEPENDENT
TARGET
GENES
EXPRESSION
.....................................
101
6.2.4
PHARMACOLOGICAL
INHIBITION
OF
STAT5
SIGNALING
AND
TARGETS
BY
+(-)
JQ1
ACTION
ON
UPSTREAM
JAK
AND
NF
K
B
MEDIATORS
IN
CLL
.......................................
104
6.2.5 TRANSCRIPTOME
ANALYSIS
AFTER
BET
INHIBITION
IN
MEC1
CLL
B-CELLS
........
106
3
6.2.6
EPIGENETIC
TARGETED
THERAPY
BY
JQ1
INHIBITOR
AFFECTS
LEUKEMIC
BURDEN
IN
CLL
TRANSPLANTED
NOG
MICE
.............................................................................
110
6.2.7
JQ1
THERAPY
INVERSES
CLL
EXPANSION
AND
IMPROVES
CYTOTOXIC
T
CELL
SUBSETS
IN
CLL
DEVELOPED
EU-TCL1
MICE
........................................................................
116
7
DISCUSSION
.............................................................................................
121
7.1
JNK1
INHIBITION
IN
CLL
.....................................................................................
121
7.2
A
MODEL
OF
JNK1
INHIBITION
IN
CLL
..................................................................
123
7.3
EFFICACY
OF
JQ1
INHIBITOR
IN
CLL
.......................................................................
125
7.4
A
MODEL
OF
BET
INHIBITION
IN
CLL
...................................................................
130
8
REFERENCES
.............................................................................................
132
9
APPENDIX
................................................................................................
149
9.1.
SUPPLEMENTARY
FIGURES
1
...............................................................................
149
9.2.
SUPPLEMENTARY
FIGURES
II
..............................................................................
153
9.3.
ABBREVIATIONS
.................................................................................................
160
9.4.
LIST
OF
FIGURES
................................................................................................
165
9.5.
LIST
OF
TABLES
.................................................................................................
167
9.6.
PUBLICATIONS
...................................................................................................
168
9.7.
CURRICULUM
VITAE
............................................................................................
169
10
ACKNOWLEDGEMENTS
...............................................................................
171
4
|
| adam_txt |
TABLE
OF
CONTENTS
1
SUMMARY
.
5
1.1
SUMMARY
I
.
5
1.2
SUMMARY
II
.
6
2
INTRODUCTION
.
7
2.1
CHRONIC
LYMPHOCYTIC
LEUKEMIA
.
7
2.1.1
CHARACTERISTICS
OF
CLL
.
7
2.1.2
CLINICAL
STAGING
AND
PROGNOSTIC
MARKERS
IN
CLL
.
7
2.2
CLL
MICROENVIRONMENT
.
9
2.3
CELL
SIGNALING
IN
CLL
.
12
2.3.1
B-CELL
RECEPTOR
PATHWAY
.
12
2.3.2
JNK1
SIGNALING
.
22
2.3.3
NOTCH
AND
NFKB
SIGNALING
.
23
2.4
THERAPEUTIC
OPTIONS
IN
B-CLL
CELL
RELATED
IMMUNOLOGICAL
DISEASES
.
24
2.4.1
SIGNALING
TARGETS
FOR
CLL
THERAPY
.
24
2.4.2 EPIGENETIC
TARGETED
THERAPIES
IN
CLL
.
31
3
AIMS
OF
THE
STUDY
.
34
4
MATERIALS
.
35
4.1
LABORATORY
EQUIPMENTS
AND
DISPOSABLES
.
35
4.1.1
EQUIPMENT
.
35
4.1.2
DISPOSABLES
.
37
4.1.3
CHEMICALS
AND
REAGENTS
.
38
4.1.4
BUFFERS
AND
SOLUTIONS
.
41
4.2
MEDIA
.
43
4.3
KITS
.
44
4.4
HUMAN
BLOOD
SAMPLES
.
44
4.4.1.
PRIMARY
HUMAN
CLL
SAMPLES
.
44
1
4.4.2.
HEALTHY
DONOR
BLOOD
SAMPLES
.
44
4.5
CELL
LINES
.
45
4.6
ANTIBODIES
.
45
4.6.1.
FACS
.
45
4.6.2.
CO-IMMUNOPRECIPITATION
AND
WESTERNBLOT
.
46
4.7
.
INHIBITORS
AND
SIRNA
.
50
4.7.1.
JNK1
AND
BROMODOMAIN
INHIBITORS
.
50
4.7.2.
SIRNAS
.
50
4.8
.
OLIGONUCLEOTIDES
.
51
4.9
.
MOUSE
STRAINS
AND
ANIMAL
HUSBANDRY
.
51
4.10
.
DATABASES
.
52
4.11
.
SOFTWARE
AND
TOOLS
.
52
5
METHODS
.
54
5.1.
IN
VITRO
EXPERIMENTS
.
54
5.1.1.
CELL
CULTURE
.
54
5.1.2.
B
CELL
ISOLATION
.
55
5.1.3.
INHIBITORS
TREATMENT
.
55
5.1.4.
SIRNA
MEDIATED
KNOCK
DOWN
.
56
5.1.5.
APOPTOSIS
ASSAY
.
56
5.1.6.
PROLIFERATION
ASSAY
.
57
5.1.7.
TRANSWELL
MIGRATION
ASSAY
.
58
5.1.8.
CXCR4
SURFACE
EXPRESSION
.
58
5.1.9.
PROTEIN
ANALYSIS
.
58
5.1.10.
CELL
CYCLE
ANALYSIS
.
60
5.1.11.
RNA
EXTRACTION,
REVERSE
TRANSCRIPTION
AND
QPCR
(TAQMAN)
.
60
5.1.12.
GENE
ARRAY
(BY
DR.
DIETMAR
PFEIFER)
.
61
5.2.
IN
VIVO
EXPERIMENTS
.
61
5.2.1
XENOGRAFT
MOUSE
MODEL
.
61
5.2.2.
EP-TCL1
MOUSE
MODEL
.
61
2
5.2.3.
TREATMENT
EXPERIMENTS
.
62
5.2.4.
BLOOD
WITHDRAWAL
AND
PROCESSING
OF
BLOOD
SAMPLES
.
63
5.2.5.
MOUSE
FOLLOW-UP
AND
SACRIFICE
THE
MICE
AFTER
DRUG
THERAPY
.
63
5.2.6.
FACS
.
64
5.2.7.
IMMUNOHISTOCHEMISTRY
(BY
DR.
KONRAD
AUMANN)
.
73
5.3.
IN
SILICO
ANALYSIS
(BY
DR.
DIETMAR
PFEIFER)
.
73
5.4.
STATISTICAL
ANALYSIS
.
73
6
RESULTS
.
74
6.1
RESULTS
1
.
74
6.1.1.
JNK1
IS
OVEREXPRESSED
AND
HYPERPHOSPHORYLATED
IN
HUMAN
PRIMARY
CLL
CELLS
74
6.1.2.
JNK1
KNOCK-DOWN
AND
JNK1
INHIBITION
AFFECTS
PRIMARY
CLL
CELL
VIABILITY.
76
6.1.3.
JNK1
INHIBITORS
REDUCE
ANTI-APOPTOTIC
DOWNSTREAM
EFFECTORS
IN
CLL
.
80
6.1.4.
SP600125
INHIBITOR
INDUCES
APOPTOSIS
AND
AFFECTS
THE
DOWNSTREAM
TARGETS
IN
UNMUTATED
CLL
IN
THE
PRESENCE
OF
PROTECTIVE
STROMAL
CELLS
.
81
6.1.5.
JNK1
IS
A
CRUCIAL
DOWNSTREAM
MEDIATOR
OF
B-CELL
RECEPTOR
SIGNALING
IN
CLL
82
6.1.6.
JNK1
INHIBITION
REDUCES
LEUKEMIC
BURDEN
IN
SPLEEN
AND
BLOOD
OF
CLL
TRANSPLANTED
NOG
MICE
.
85
6.1.7.
JNK1
INHIBITOR
DECREASES
LEUKEMIC
CLL
CELLS
AND
MODULATES
IMMUNE
RESPONSES
IN
EP-TCL1
MOUSE
MODEL
.
89
6.2
RESULTS
II
.
94
6.2.1
FUNCTIONAL
ANALYSIS
AFTER
BET
INHIBITION
IN
B-CLL
CELL
LINE
AND
PRIMARY
CLL
CELLS
94
6.2.2
BET
INHIBITOR
AFFECTS
STAT5
AND
OTHER
PROLIFERATIVE
PROTEINS
INVOLVE
IN
CLL
CELL
PROLIFERATION
AND
SURVIVAL
.
98
6.2.3
JQ1
AFFECTS
MIGRATORY
EFFECT
OF
CLL
CELLS
THROUGH
REGULATION
OF
STAT5
DEPENDENT
AND
INDEPENDENT
TARGET
GENES
EXPRESSION
.
101
6.2.4
PHARMACOLOGICAL
INHIBITION
OF
STAT5
SIGNALING
AND
TARGETS
BY
+(-)
JQ1
ACTION
ON
UPSTREAM
JAK
AND
NF
K
B
MEDIATORS
IN
CLL
.
104
6.2.5 TRANSCRIPTOME
ANALYSIS
AFTER
BET
INHIBITION
IN
MEC1
CLL
B-CELLS
.
106
3
6.2.6
EPIGENETIC
TARGETED
THERAPY
BY
JQ1
INHIBITOR
AFFECTS
LEUKEMIC
BURDEN
IN
CLL
TRANSPLANTED
NOG
MICE
.
110
6.2.7
JQ1
THERAPY
INVERSES
CLL
EXPANSION
AND
IMPROVES
CYTOTOXIC
T
CELL
SUBSETS
IN
CLL
DEVELOPED
EU-TCL1
MICE
.
116
7
DISCUSSION
.
121
7.1
JNK1
INHIBITION
IN
CLL
.
121
7.2
A
MODEL
OF
JNK1
INHIBITION
IN
CLL
.
123
7.3
EFFICACY
OF
JQ1
INHIBITOR
IN
CLL
.
125
7.4
A
MODEL
OF
BET
INHIBITION
IN
CLL
.
130
8
REFERENCES
.
132
9
APPENDIX
.
149
9.1.
SUPPLEMENTARY
FIGURES
1
.
149
9.2.
SUPPLEMENTARY
FIGURES
II
.
153
9.3.
ABBREVIATIONS
.
160
9.4.
LIST
OF
FIGURES
.
165
9.5.
LIST
OF
TABLES
.
167
9.6.
PUBLICATIONS
.
168
9.7.
CURRICULUM
VITAE
.
169
10
ACKNOWLEDGEMENTS
.
171
4 |
| any_adam_object | 1 |
| any_adam_object_boolean | 1 |
| author | Khaja Saleem, Al Ahmirathus Syedush Shifa 1990- |
| author_GND | (DE-588)126068105X |
| author_facet | Khaja Saleem, Al Ahmirathus Syedush Shifa 1990- |
| author_role | aut |
| author_sort | Khaja Saleem, Al Ahmirathus Syedush Shifa 1990- |
| author_variant | s a a s s k saass saassk |
| building | Verbundindex |
| bvnumber | BV049304362 |
| ctrlnum | (OCoLC)1331712623 (DE-599)KXP1807548406 |
| discipline | Medizin |
| discipline_str_mv | Medizin |
| format | Thesis Book |
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| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV049304362 |
| illustrated | Illustrated |
| index_date | 2024-07-03T22:39:34Z |
| indexdate | 2024-07-10T10:01:02Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-034565569 |
| oclc_num | 1331712623 |
| open_access_boolean | |
| owner | DE-355 DE-BY-UBR |
| owner_facet | DE-355 DE-BY-UBR |
| physical | 171 Seiten Illustrationen, Diagramme 30 cm |
| publishDate | 2021 |
| publishDateSearch | 2021 |
| publishDateSort | 2021 |
| record_format | marc |
| spelling | Khaja Saleem, Al Ahmirathus Syedush Shifa 1990- Verfasser (DE-588)126068105X aut Innovative therapeutic approaches in chronic lymphocytic leukemia vorgelegt von Al Ahmirathus Syedush Shifa Khaja Saleem Freiburg im Breisgau Oktober 2021 171 Seiten Illustrationen, Diagramme 30 cm txt rdacontent n rdamedia nc rdacarrier Dissertation Albert-Ludwigs-Universität Freiburg im Breisgau 2022 (DE-588)4113937-9 Hochschulschrift gnd-content B:DE-101 application/pdf https://d-nb.info/1264509839/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=034565569&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis 1\p emakn 0,83402 20220812 DE-101 https://d-nb.info/provenance/plan#emakn 2\p dnb 20220909 DE-101 https://d-nb.info/provenance/plan#dnb |
| spellingShingle | Khaja Saleem, Al Ahmirathus Syedush Shifa 1990- Innovative therapeutic approaches in chronic lymphocytic leukemia |
| subject_GND | (DE-588)4113937-9 |
| title | Innovative therapeutic approaches in chronic lymphocytic leukemia |
| title_auth | Innovative therapeutic approaches in chronic lymphocytic leukemia |
| title_exact_search | Innovative therapeutic approaches in chronic lymphocytic leukemia |
| title_exact_search_txtP | Innovative therapeutic approaches in chronic lymphocytic leukemia |
| title_full | Innovative therapeutic approaches in chronic lymphocytic leukemia vorgelegt von Al Ahmirathus Syedush Shifa Khaja Saleem |
| title_fullStr | Innovative therapeutic approaches in chronic lymphocytic leukemia vorgelegt von Al Ahmirathus Syedush Shifa Khaja Saleem |
| title_full_unstemmed | Innovative therapeutic approaches in chronic lymphocytic leukemia vorgelegt von Al Ahmirathus Syedush Shifa Khaja Saleem |
| title_short | Innovative therapeutic approaches in chronic lymphocytic leukemia |
| title_sort | innovative therapeutic approaches in chronic lymphocytic leukemia |
| topic_facet | Hochschulschrift |
| url | https://d-nb.info/1264509839/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=034565569&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT khajasaleemalahmirathussyedushshifa innovativetherapeuticapproachesinchroniclymphocyticleukemia |
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