Establishment and evaluation of single-cell RNA-sequencing methods to investigate tumor heterogeneity in newly generated pediatric glioma models:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
Heidelberg
2023
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis Inhaltsverzeichnis |
| Beschreibung: | x, 114 Seiten Illustrationen, Diagramme 30 cm 1 CD-ROM |
Internformat
MARC
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| 100 | 1 | |a Ernst, Kati |d 1990- |e Verfasser |0 (DE-588)1317559827 |4 aut | |
| 245 | 1 | 0 | |a Establishment and evaluation of single-cell RNA-sequencing methods to investigate tumor heterogeneity in newly generated pediatric glioma models |c presented by Kati Johanna Ernst née Lappalainen, M. Sc. |
| 264 | 1 | |a Heidelberg |c 2023 | |
| 300 | |a x, 114 Seiten |b Illustrationen, Diagramme |c 30 cm |e 1 CD-ROM | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |b Dissertation |c Ruprecht-Karls-University Heidelberg |d 2023 | ||
| 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/1316908615/04 |3 Inhaltsverzeichnis |
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Datensatz im Suchindex
| _version_ | 1823147005725114368 |
|---|---|
| adam_text |
CONTENT
ABBREVIATIONS
.
III
SUMMARY
.
VII
ZUSAMMENFASSUNG
.
IX
1
INTRODUCTION
.
1
1.1
PEDIATRIC
CANCER
.
1
1.1.1
PEDIATRIC
GLIOMA
.
2
1.1.2
CURRENT
THERAPIES
FOR
PEDIATRIC
GLIOMA
.
7
1.2
PRECLINICAL
MOUSE
MODELS
.
8
1.2.1
TRANSPLANT-BASED
AND
GENETICALLY
ENGINEERED
MOUSE
MODELS
.
9
1.2.1.1
TRANSPLANT-BASED
MODELS
.
9
1.2.1.2
MODELS
WITH
GERMLINE
ALTERATION
.
10
1.2.1.4
MODELS
WITH
SOMATIC
ALTERATION
.
12
1.3
SINGLE-CELL
SEQUENCING
.
14
1.3.1
HISTORY
OF
SINGLE-CELL
SEQUENCING
.
14
1.3.2
TUMOR
HETEROGENEITY
.
16
1.3.2.1
TUMOR
HETEROGENEITY
OF
PHGG
.
18
1.4
AIMS
OF
THIS
THESIS
.
21
2
MATERIALS
&
METHODS
.
22
2.1
MATERIALS
.
22
2.1.1
CHEMICALS,
REAGENTS
AND
FURTHER
MATERIAL
.
22
2.1.2
ENZYMES
.
23
2.1.3
ANTIBODIES
.
23
2.1.4
PHARMACEUTICAL
DRUGS
AND
SUBSTANCES
.
23
2.1.5
OLIGONUCLEOTIDES
.
24
2.1.6
PLASMIDS
.
24
2.1.7
MOUSE
LINES
.
24
2.1.8
KITS
.
25
2.1.9
EQUIPMENT
.
25
2.1.10
INSTRUMENTS
.
26
2.1.11
SOFTWARE
.
26
2.2
METHODS
.
27
2.2.1
INTRACRANIAL
INJECTION
.
27
2.2.2
IN-UTERO
ELECTROPORATION
.
28
2.2.3
LUCIFERASE
IMAGING
.
30
2.2.4
HISTOLOGICAL
ANALYSIS
.
30
2.2.5
PRIMARY
TUMOR
CELL
CULTURE
.
32
2.2.6
SEQUENCING
OF
THE
CRISPR
TARGET
LOCI
.
33
2.2.7
QUANTITATIVE
PCR
.
33
2.2.8
OPTIMIZATION
OF
NUCLEAR
EXTRACTION
FROM
FROZEN
TISSUES
.
34
2.2.9
COMPARISON
TO
OTHER
NUCLEAR
EXTRACTION
METHODS
.
35
2.2.10
NUCLEI
STAINING
EXPERIMENTS
.
35
2.2.11
TRANSCRIPTOME
AMPLIFICATION
AND
CDNA
LIBRARY
PREPARATION
.
36
2.2.12
NEXT
GENERATION
SEQUENCING
.
36
2.2.13
COMPUTATIONAL
DATA
ANALYSIS
.
37
2.2.13.1
SNRNA-SEQ
METHOD
COMPARISON
.37
2.2.13.2
COMPARISON
OF
PRECLINICAL
MOUSE
MODELS
.38
2.2.14
ETHICAL
JUSTIFICATION
.
39
3
RESULTS
.
40
3.1
SOMATIC
MOUSE
MODELS
FOR
PHGG
.
40
3.2
STUDY
OF
TUMOR GROWTH
PATTERN
.
45
3.3
ESTABLISHMENT
OF
A
SINGLE-NUCLEUS
RNA-SEQ
METHOD
.
48
3.3.1
ISOLATION
OF
INTACT
NUCLEI
FROM
FROZEN
TISSUES
.
49
3.3.2
COMPARISON
OF
SCRNA-SEQ
PLATFORMS
USING
A
PDX
MODEL
.
51
3.3.3
SNRNA-SEQ
OF
FROZEN
PRIMARY
TUMOR
TISSUES
.
58
3.3.4
CNV
ANALYSIS
SUPPORTS
TUMOR
CELL
DETECTION
FROM
10X
DATA
.
65
3.4
INVESTIGATING
TUMOR
HETEROGENEITY
OF
PRECLINICAL
MOUSE
MODELS
.
68
3.4.1
DISTINCTIVE
FEATURES
OF
PHGG
MOUSE
MODELS
.
68
3.4.1.1
.
SAMPLE
COHORT
.
69
3.4.1.2
DIFFERENT
CELL
TYPE
DISTRIBUTION
BETWEEN
H3-K27M
AND
MET-FUSION
ALTERED
PHGG
.
72
3.4.1.3
A
TUMOR
STATE
CLOSEST
TO
THE
MALIGNANT
CELL-OF-ORIGIN
SUPPORTED
BY
TAMS
WAS
ENRICHED
IN
THE
PDXS
73
3.4.1.4
ALL
CELL
POPULATIONS
WERE
RETAINED
IN
ALLOGRAFTS
.
75
3.4.2
THE
IMMUNE
SYSTEM
POSSIBLY
LIMITS
THE
TUMOR
GROWTH
THREE
WEEKS
AFTER
TUMOR
INDUCTION
IN
MICE
78
3.4.3
CELL
POPULATIONS
LOCATE
DIFFERENTLY
WITHIN
A
TUMOR
.
80
4
DISCUSSION
AND
OUTLOOK
.
83
4.1
EVALUATION
OF
PHGG MODEL
GENERATION
.
83
4.1.1
EFFECT
OF
GENETIC
ALTERATIONS
.
83
4.1.2
LOCATION
AND
GROWTH
PATTERN
.86
4.2
NUCLEUS
ISOLATION
AND
SNRNA-SEQ
METHOD
.
88
4.3
TUMOR
HETEROGENEITY
OF
PRECLINICAL PHGG
MODELS
.
89
4.3.1
CELL
COMPOSITION
IN
ALLOGRAFTS
AND
XENOGRAFTS
.
89
4.3.2
INVOLVEMENT
OF
IMMUNE
SYSTEM
.
92
4.4
CONCLUDING
REMARKS
.
95
5
REFERENCES
.
96
6
PUBLICATIONS
.
108
7
APPENDIX
.
110
8
ACKNOWLEDGEMENTS
.
112 |
| any_adam_object | 1 |
| author | Ernst, Kati 1990- |
| author_GND | (DE-588)1317559827 |
| author_facet | Ernst, Kati 1990- |
| author_role | aut |
| author_sort | Ernst, Kati 1990- |
| author_variant | k e ke |
| building | Verbundindex |
| bvnumber | BV050152838 |
| ctrlnum | (DE-599)DNB1316908615 |
| discipline | Medizin |
| format | Thesis Book |
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| id | DE-604.BV050152838 |
| illustrated | Illustrated |
| indexdate | 2025-02-04T17:01:14Z |
| institution | BVB |
| language | English |
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| owner | DE-355 DE-BY-UBR |
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| physical | x, 114 Seiten Illustrationen, Diagramme 30 cm 1 CD-ROM |
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| publishDateSearch | 2023 |
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| spelling | Ernst, Kati 1990- Verfasser (DE-588)1317559827 aut Establishment and evaluation of single-cell RNA-sequencing methods to investigate tumor heterogeneity in newly generated pediatric glioma models presented by Kati Johanna Ernst née Lappalainen, M. Sc. Heidelberg 2023 x, 114 Seiten Illustrationen, Diagramme 30 cm 1 CD-ROM txt rdacontent n rdamedia nc rdacarrier Dissertation Ruprecht-Karls-University Heidelberg 2023 (DE-588)4113937-9 Hochschulschrift gnd-content B:DE-101 application/pdf https://d-nb.info/1316908615/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=035489117&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis 1\p emakn 0,59317 20240206 DE-101 https://d-nb.info/provenance/plan#emakn 2\p dnb 20240220 DE-101 https://d-nb.info/provenance/plan#dnb 3\p dnb 20240220 DE-101 https://d-nb.info/provenance/plan#dnb |
| spellingShingle | Ernst, Kati 1990- Establishment and evaluation of single-cell RNA-sequencing methods to investigate tumor heterogeneity in newly generated pediatric glioma models |
| subject_GND | (DE-588)4113937-9 |
| title | Establishment and evaluation of single-cell RNA-sequencing methods to investigate tumor heterogeneity in newly generated pediatric glioma models |
| title_auth | Establishment and evaluation of single-cell RNA-sequencing methods to investigate tumor heterogeneity in newly generated pediatric glioma models |
| title_exact_search | Establishment and evaluation of single-cell RNA-sequencing methods to investigate tumor heterogeneity in newly generated pediatric glioma models |
| title_full | Establishment and evaluation of single-cell RNA-sequencing methods to investigate tumor heterogeneity in newly generated pediatric glioma models presented by Kati Johanna Ernst née Lappalainen, M. Sc. |
| title_fullStr | Establishment and evaluation of single-cell RNA-sequencing methods to investigate tumor heterogeneity in newly generated pediatric glioma models presented by Kati Johanna Ernst née Lappalainen, M. Sc. |
| title_full_unstemmed | Establishment and evaluation of single-cell RNA-sequencing methods to investigate tumor heterogeneity in newly generated pediatric glioma models presented by Kati Johanna Ernst née Lappalainen, M. Sc. |
| title_short | Establishment and evaluation of single-cell RNA-sequencing methods to investigate tumor heterogeneity in newly generated pediatric glioma models |
| title_sort | establishment and evaluation of single cell rna sequencing methods to investigate tumor heterogeneity in newly generated pediatric glioma models |
| topic_facet | Hochschulschrift |
| url | https://d-nb.info/1316908615/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=035489117&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT ernstkati establishmentandevaluationofsinglecellrnasequencingmethodstoinvestigatetumorheterogeneityinnewlygeneratedpediatricgliomamodels |
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