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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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Related in: MedlinePlus

Tumor histology.H & E sections of (A) HROC40-PDX and (B) its primary. Note the invasive edge towards the right. Principal morphological features are retained in the PDX. β-catenin immunohistochemistry of (C) HROC60-PDX and (D) its primary. Note nuclear β-catenin translocation at the invasive edge of both tumors.
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pone.0143194.g001: Tumor histology.H & E sections of (A) HROC40-PDX and (B) its primary. Note the invasive edge towards the right. Principal morphological features are retained in the PDX. β-catenin immunohistochemistry of (C) HROC60-PDX and (D) its primary. Note nuclear β-catenin translocation at the invasive edge of both tumors.

Mentions: Surface marker expression was done by flow cytometry with and without IFN-γ (200 IU/mL for 48 hours) pre-treatment using a panel of Abs (for details see Fig 1B). Samples were analyzed using CellQuest software (BD Biosciences). Additionally, multi-color flow cytometry was done on a FACSAria using following mAbs: anti-CD47-BV421, anti-CD36-PE, anti-CD73-BV510, anti-CD95-FITC, anti-Foxp3-Alexa Fluor 647, anti-CD284-BV421, anti-CD276-PE, anti-CD133-APC, anti-cFLIP-Alexa Fluor 488, anti-Indolamin-2.3-Dioxygenase-PE, anti-BIN1 (bridging integrator 1)-Alexa Fluor 488.


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)

Tumor histology.H & E sections of (A) HROC40-PDX and (B) its primary. Note the invasive edge towards the right. Principal morphological features are retained in the PDX. β-catenin immunohistochemistry of (C) HROC60-PDX and (D) its primary. Note nuclear β-catenin translocation at the invasive edge of both tumors.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0143194.g001: Tumor histology.H & E sections of (A) HROC40-PDX and (B) its primary. Note the invasive edge towards the right. Principal morphological features are retained in the PDX. β-catenin immunohistochemistry of (C) HROC60-PDX and (D) its primary. Note nuclear β-catenin translocation at the invasive edge of both tumors.
Mentions: Surface marker expression was done by flow cytometry with and without IFN-γ (200 IU/mL for 48 hours) pre-treatment using a panel of Abs (for details see Fig 1B). Samples were analyzed using CellQuest software (BD Biosciences). Additionally, multi-color flow cytometry was done on a FACSAria using following mAbs: anti-CD47-BV421, anti-CD36-PE, anti-CD73-BV510, anti-CD95-FITC, anti-Foxp3-Alexa Fluor 647, anti-CD284-BV421, anti-CD276-PE, anti-CD133-APC, anti-cFLIP-Alexa Fluor 488, anti-Indolamin-2.3-Dioxygenase-PE, anti-BIN1 (bridging integrator 1)-Alexa Fluor 488.

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