Imaging of tumor clones with differential liver colonization

Oshima, Go; Wightman, Sean C.; Uppal, Abhineet; Stack, Melinda E.; Pitroda, Sean P.; Oskvarek, Jonathan J.; Huang, Xiaona; Posner, Mitchell C.; Hellman, Samuel; Weichselbaum, Ralph R.; Khodarev, Nikolai N.

doi:10.1038/srep10946

Cited by 9 publications

(20 citation statements)

References 34 publications

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“…7). To further explore the impact of DNA demethylation and 14q32 miRNA expression on metastatic development, we quantified metastatic colonization of the liver by WT and DKO cells in athymic nude mice (18,19). As expected, DNA demethylation as a result of DNMT1/3B disruption completely abrogated metastatic colonization of DKO cells ( Supplementary Fig.…”

Section: Overexpression Of Dna Methylation-dependent 14q32 Mirnas In mentioning

confidence: 68%

“…The generation of colorectal liver metastases and quantification of metastatic burden were performed as described previously (19). Briefly, HCT116-derived cells were injected into the spleens of anesthetized athymic nude mice (5-6 weeks old; Envigo), followed by complete splenectomy.…”

Section: Murine Model Of Colorectal Liver Metastasismentioning

confidence: 99%

“…The model of colorectal liver metastasis exploited in this report provides a quantitative measure of the number of metastatic colonies, which is proportional to colonization ability of metastatic clones, and size of individual colonies, which is proportional to the growth propensity of corresponding clones. We recently demonstrated that these parameters are relatively independent and may reflect different mechanisms of metastatic propensity (19). In addition, colonization frequency at least partially depends on the adhesion, invasion, and migration properties of metastatic clones (2, 6).…”

Section: Reactivation Of 14q32 Mirnas Is Sufficient To Restrict Livermentioning

confidence: 99%

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DNA Methylation Controls Metastasis-Suppressive 14q32-Encoded miRNAs

et al. 2019

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Expression of 14q32-encoded miRNAs is a favorable prognostic factor in patients with metastatic cancer. In this study, we used genomic inhibition of DNA methylation through disruption of DNA methyltransferases DNMT1 and DNMT3B and pharmacologic inhibition with 5-Aza-2 0-deoxycytidine (5-Aza-dC, decitabine) to demonstrate that DNA methylation predominantly regulates expression of metastasis-suppressive miRNAs in the 14q32 cluster. DNA demethylation facilitated CCCTC-binding factor (CTCF) recruitment to the maternally expressed gene 3 differentially methylated region (MEG3-DMR), which acts as a cis-regulatory element for 14q32 miRNA expression. 5-Aza-dC activated demethylation of the MEG3-DMR and expression of 14q32 miRNAs, which suppressed adhesion, invasion, and migration (AIM) properties of met-astatic tumor cells. Cancer cells with MEG3-DMR hypomethylation exhibited constitutive expression of 14q32 miRNAs and resistance to 5-Aza-dC-induced suppression of AIM. Expression of methylation-dependent 14q32 miRNAs suppressed metastatic colonization in preclinical models of lung and liver metastasis and correlated with improved clinical outcomes in patients with metastatic cancer. These findings implicate epigenetic modification via DNA methylation in the regulation of metastatic propensity through miRNA networks and identify a previously unrecognized action of decitabine on the activation of metastasis-suppressive miRNAs. Significance: This study investigates epigenetic regulation of metastasis-suppressive miRNAs and the effect on metastasis.

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Section: Overexpression Of Dna Methylation-dependent 14q32 Mirnas In mentioning

confidence: 68%

Section: Murine Model Of Colorectal Liver Metastasismentioning

confidence: 99%

Section: Reactivation Of 14q32 Mirnas Is Sufficient To Restrict Livermentioning

confidence: 99%

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DNA Methylation Controls Metastasis-Suppressive 14q32-Encoded miRNAs

et al. 2019

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“…First, in order to ensure the ability to observe metastatic clones with different propensities to colonize and proliferate in the liver, a panel of highly heterogeneous monoclonal cell lines was established, rather than an established unfractionated cancer cell line 12,13 . The monoclonal approach to metastasis development is justified by recent genomic data 14 and was successfully used previously to model the metastatic process 10,15,16 . Second, double-labeling of the parental cell line by luciferase and tdTomato markers was incorporated in order to provide enhanced analysis of metastatic growth both in vivo and ex vivo.…”

Section: Discussionmentioning

confidence: 99%

Advanced Animal Model of Colorectal Metastasis in Liver: Imaging Techniques and Properties of Metastatic Clones

Oshima

Stack

Wightman

et al. 2016

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Patients with a limited number of hepatic metastases and slow rates of progression can be successfully treated with local treatment approaches. However, little is known about the heterogeneity of liver metastases, and animal models capable of evaluating the development of individual metastatic colonies are needed. Here, we present an advanced model of hepatic metastases that provides the ability to quantitatively visualize the development of individual tumor clones in the liver and estimate their growth kinetics and colonization efficiency. We generated a panel of monoclonal derivatives of HCT116 human colorectal cancer cells stably labeled with luciferase and tdTomato and possessing different growth properties. With a splenic injection followed by a splenectomy, the majority of these clones are able to generate hepatic metastases, but with different frequencies of colonization and varying growth rates. Using the In Vivo Imaging System (IVIS), it is possible to visualize and quantify metastasis development with in vivo luminescent and ex vivo fluorescent imaging. In addition, Diffuse Luminescent Imaging Tomography (DLIT) provides a 3D distribution of liver metastases in vivo. Ex vivo fluorescent imaging of harvested livers provides quantitative measurements of individual hepatic metastatic colonies, allowing for the evaluation of the frequency of liver colonization and the growth kinetics of metastases. Since the model is similar to clinically observed liver metastases, it can serve as a modality for detecting genes associated with liver metastasis and for testing potential ablative or adjuvant treatments for liver metastatic disease.

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“…Liver metastases of colorectal cancer were established as previously described. 68 Briefly, 2 × 10 6 HCT116-L2T cells stably transfected with luciferin and tdTomato were slowly injected into the spleens of athymic nude mice, respectively, anesthetized with 2% isoflurane in oxygen (v/v). The spleens were removed 5 min later to reduce tumor growth in the spleen and the abdominal cavity.…”

Section: Establishment Of a Xenogenic Hepatic Metastasis Tumor Modelmentioning

confidence: 99%

Systemic miRNA delivery by nontoxic nanoscale coordination polymers limits epithelial-to-mesenchymal transition and suppresses liver metastases of colorectal cancer

et al. 2019

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Though early detection and treatment of primary tumors has significantly improved in recent years, metastatic disease remains among the most significant challenges in cancer therapy. Cancer cells can disseminate before the primary tumor is detected to form micro or gross metastases, requiring toxic systemic therapies. To prevent and suppress metastases, we have developed a nontoxic, long-circulating nanoscale coordination polymer (NCP) protecting microRNA (miRNA) in circulation and releasing it in tumors. Pt IV (en) 2 [en=ethylenediamine] containing NCPs (PtEN) can release a nontoxic, kinetically inert Pt II (en) 2 compound and carbon dioxide which aids the endosomal escape of its miRNA cargo, miR-655-3p. Without the presence of the PtEN core, the miRNA showed cellular uptake but no effect. When transfected into human colorectal HCT116 cells by NCPs, this oligometastatic miRNA limited proliferation and epithelial-to-mesenchymal transition (EMT) by preventing β-catenin nuclear translocation and tumor cell invasion. Systemic administrations of PtEN/miR-655-3p sustained effective transfection to reduce liver colonization and tumor burden in a xenogenic hepatic metastatic model of HCT116 without any observable toxicity.

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Imaging of tumor clones with differential liver colonization

Cited by 9 publications

References 34 publications

DNA Methylation Controls Metastasis-Suppressive 14q32-Encoded miRNAs

DNA Methylation Controls Metastasis-Suppressive 14q32-Encoded miRNAs

Advanced Animal Model of Colorectal Metastasis in Liver: Imaging Techniques and Properties of Metastatic Clones

Systemic miRNA delivery by nontoxic nanoscale coordination polymers limits epithelial-to-mesenchymal transition and suppresses liver metastases of colorectal cancer

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