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Functional Characterization and Drug Response of Freshly Established Patient-Derived Tumor Models with CpG Island Methylator Phenotype.

Maletzki C, Huehns M, Knapp P, Waukosin N, Klar E, Prall F, Linnebacher M - PLoS ONE (2015)

Bottom Line: Initial DNA fingerprint analysis confirmed identity with the patient in all four cases.These freshly established cells showed characteristic features associated with the CIMP-phenotype (HROC40: APCwt, TP53 mut, KRAS mut; 3/8 marker methylated; HROC43: APC mut, TP53 mut, KRAS mut; 4/8 marker methylated; HROC60: APCwt, TP53 mut, KRASwt; 4/8 marker methylated; HROC183: APC mut, TP53 mut, KRAS mut; 6/8 marker methylated).Of note, most cell lines were sensitive towards "non-classical" CRC standard drugs (sensitivity: Gemcitabin > Rapamycin > Nilotinib).

View Article: PubMed Central - PubMed

Affiliation: Molecular Oncology and Immunotherapy, University of Rostock, Rostock, Germany.

ABSTRACT
Patient-individual tumor models constitute a powerful platform for basic and translational analyses both in vitro and in vivo. However, due to the labor-intensive and highly time-consuming process, only few well-characterized patient-derived cell lines and/or corresponding xenografts exist. In this study, we describe successful generation and functional analysis of novel tumor models from patients with sporadic primary colorectal carcinomas (CRC) showing CpG island methylator phenotype (CIMP). Initial DNA fingerprint analysis confirmed identity with the patient in all four cases. These freshly established cells showed characteristic features associated with the CIMP-phenotype (HROC40: APCwt, TP53 mut, KRAS mut; 3/8 marker methylated; HROC43: APC mut, TP53 mut, KRAS mut; 4/8 marker methylated; HROC60: APCwt, TP53 mut, KRASwt; 4/8 marker methylated; HROC183: APC mut, TP53 mut, KRAS mut; 6/8 marker methylated). Cell lines were of epithelial origin (EpCAM+) with distinct morphology and growth kinetics. Response to chemotherapeutics was quite individual between cells, with stage I-derived cell line HROC60 being most susceptible towards standard clinically approved chemotherapeutics (e.g. 5-FU, Irinotecan). Of note, most cell lines were sensitive towards "non-classical" CRC standard drugs (sensitivity: Gemcitabin > Rapamycin > Nilotinib). This comprehensive analysis of tumor biology, genetic alterations and assessment of chemosensitivity towards a broad range of (chemo-) therapeutics helps bringing forward the concept of personalized tumor therapy.

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Immunosuppressive phenotype of CIMP+ cell lines.Assessment of immune evasion markers was conducted by multi-color flow cytometry using fluorochrom-labeled mAbs as given on the x-axis. Results are given as the mean % of positive cells + standard deviation of three independent experiments.
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pone.0143194.g003: Immunosuppressive phenotype of CIMP+ cell lines.Assessment of immune evasion markers was conducted by multi-color flow cytometry using fluorochrom-labeled mAbs as given on the x-axis. Results are given as the mean % of positive cells + standard deviation of three independent experiments.

Mentions: Additional flow cytometric phenotyping confirmed their epithelial origin (>90% EpCAM+). Detailed information on cellular phenotype is given in Fig 2B and 2C. In line with their immunosuppressive phenotype, several molecules, known to be linked to cancer progression and immune evasion, were highly expressed (>80%: CD47, CD274, CD276, and Indoleamine 2,3 dioxygenase-1). By contrast, an individual profile was seen for cFLIP (cellular FLICE (FADD-like IL-1β-converting enzyme)-inhibitory protein), CD73, CD95 and CD133 (Fig 3).


Functional Characterization and Drug Response of Freshly Established Patient-Derived Tumor Models with CpG Island Methylator Phenotype.

Maletzki C, Huehns M, Knapp P, Waukosin N, Klar E, Prall F, Linnebacher M - PLoS ONE (2015)

Immunosuppressive phenotype of CIMP+ cell lines.Assessment of immune evasion markers was conducted by multi-color flow cytometry using fluorochrom-labeled mAbs as given on the x-axis. Results are given as the mean % of positive cells + standard deviation of three independent experiments.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4664421&req=5

pone.0143194.g003: Immunosuppressive phenotype of CIMP+ cell lines.Assessment of immune evasion markers was conducted by multi-color flow cytometry using fluorochrom-labeled mAbs as given on the x-axis. Results are given as the mean % of positive cells + standard deviation of three independent experiments.
Mentions: Additional flow cytometric phenotyping confirmed their epithelial origin (>90% EpCAM+). Detailed information on cellular phenotype is given in Fig 2B and 2C. In line with their immunosuppressive phenotype, several molecules, known to be linked to cancer progression and immune evasion, were highly expressed (>80%: CD47, CD274, CD276, and Indoleamine 2,3 dioxygenase-1). By contrast, an individual profile was seen for cFLIP (cellular FLICE (FADD-like IL-1β-converting enzyme)-inhibitory protein), CD73, CD95 and CD133 (Fig 3).

Bottom Line: Initial DNA fingerprint analysis confirmed identity with the patient in all four cases.These freshly established cells showed characteristic features associated with the CIMP-phenotype (HROC40: APCwt, TP53 mut, KRAS mut; 3/8 marker methylated; HROC43: APC mut, TP53 mut, KRAS mut; 4/8 marker methylated; HROC60: APCwt, TP53 mut, KRASwt; 4/8 marker methylated; HROC183: APC mut, TP53 mut, KRAS mut; 6/8 marker methylated).Of note, most cell lines were sensitive towards "non-classical" CRC standard drugs (sensitivity: Gemcitabin > Rapamycin > Nilotinib).

View Article: PubMed Central - PubMed

Affiliation: Molecular Oncology and Immunotherapy, University of Rostock, Rostock, Germany.

ABSTRACT
Patient-individual tumor models constitute a powerful platform for basic and translational analyses both in vitro and in vivo. However, due to the labor-intensive and highly time-consuming process, only few well-characterized patient-derived cell lines and/or corresponding xenografts exist. In this study, we describe successful generation and functional analysis of novel tumor models from patients with sporadic primary colorectal carcinomas (CRC) showing CpG island methylator phenotype (CIMP). Initial DNA fingerprint analysis confirmed identity with the patient in all four cases. These freshly established cells showed characteristic features associated with the CIMP-phenotype (HROC40: APCwt, TP53 mut, KRAS mut; 3/8 marker methylated; HROC43: APC mut, TP53 mut, KRAS mut; 4/8 marker methylated; HROC60: APCwt, TP53 mut, KRASwt; 4/8 marker methylated; HROC183: APC mut, TP53 mut, KRAS mut; 6/8 marker methylated). Cell lines were of epithelial origin (EpCAM+) with distinct morphology and growth kinetics. Response to chemotherapeutics was quite individual between cells, with stage I-derived cell line HROC60 being most susceptible towards standard clinically approved chemotherapeutics (e.g. 5-FU, Irinotecan). Of note, most cell lines were sensitive towards "non-classical" CRC standard drugs (sensitivity: Gemcitabin > Rapamycin > Nilotinib). This comprehensive analysis of tumor biology, genetic alterations and assessment of chemosensitivity towards a broad range of (chemo-) therapeutics helps bringing forward the concept of personalized tumor therapy.

Show MeSH
Related in: MedlinePlus