High-sensitivity Detection of Micrometastases Generated by GFP Lentivirus-transduced Organoids Cultured from a Patient-derived Colon Tumor

Okazawa, Yu; Mizukoshi, Kosuke; Koyama, Yu; Okubo, Shoki; Komiyama, Hiromitsu; Kojima, Yutaka; Goto, Michitoshi; Habu, Sonoko; Hino, Okio; Sakamoto, Kazuhiro; Orimo, Akira

doi:10.3791/57374

Cited by 7 publications

(13 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Fresh tumor specimens surgically dissected from CRC patients were implanted subcutaneously into NOG mice, as previously described . Tumor xenografts that developed, to ~1 cm 3 in size, were also resected and grossly necrotic areas were removed.…”

Section: Methodsmentioning

confidence: 99%

“…Tumor cell organoids were prepared from CRC PDXs prior to in vitro culture, as described previously . These tumor organoids were also infected with or without lentiviral particles expressing GFP and different shRNAs.…”

Section: Methodsmentioning

confidence: 99%

“…Fresh tumor specimens surgically dissected from CRC patients were implanted subcutaneously into NOG mice, as previously described. 29 Tumor xenografts that developed, to~1 cm 3 in size, were also resected and grossly necrotic areas were removed. The tumors were then minced mechanically with a razor blade and transferred into a 15 ml tube containing DMEM/F-12 with GlutaMAX™ supplemented (Thermo Fisher Scientific, Waltham, MA) with 5% fetal bovine serum (FBS) (Sigma-Aldrich, St. Louis, MO), penicillin (100 U/ml)-streptomycin (100 μg/ml, Thermo Fisher Scientific) and collagenase Type I (1.5 mg/ml; Sigma-Aldrich) followed by slow agitation for 2 hr at 37 C. The resulting solution with enzymatically digested tissues was filtrated twice using a cell strainer with a 40 μm pore size (Corning, NY).…”

Section: Establishment Of the Crc Pdx Mouse Models By Subcutaneous Anmentioning

confidence: 99%

“…Tumor cell organoids were prepared from CRC PDXs prior to in vitro culture, as described previously. 29 These tumor organoids were also infected with or without lentiviral particles expressing GFP and different shRNAs. To generate primary tumors, the CRC organoid cell suspension (50 μl) including 5 × 10 5 cells with 50% Matrigel (Corning, NY) was injected orthotopically into the rectal submucosa of NOG mice using a Hamilton syringe (needle size: 22 gauge, Tokyo Science) (Figs.…”

Section: Establishment Of Experimental Tumor and Metastasis Models Ofmentioning

confidence: 99%

“…S2b-S2d and S6a), as described previously. 29 To generate liver metastases, the CRC organoid cell suspension (50 μl) including 4 × 10 4 cells in phosphate buffered saline (PBS) was injected intrasplenically into NOG mice using a Hamilton syringe (needle size: 22 gauge, Tokyo Science) (Figs. 2f-2h, 4b, 4c, 5c, and 5d; Supplementary Figs.…”

Section: Establishment Of Experimental Tumor and Metastasis Models Ofmentioning

confidence: 99%

See 4 more Smart Citations

Metastatic seeding of human colon cancer cell clusters expressing the hybrid epithelial/mesenchymal state

Mizukoshi

Okazawa

Haeno

et al. 2019

Intl Journal of Cancer

Self Cite

View full text Add to dashboard Cite

Emerging evidence supports the theory that tumor cell clusters efficiently metastasize to distant organs. However, the roles of epithelial‐to‐mesenchymal transition (EMT) in metastasizing tumor cell clusters have not yet been fully elucidated. To investigate this issue, tumor fragments were dissected from 40 colorectal cancer (CRC) patients and implanted subcutaneously into immunodeficient mice. We observed that tumors developed from the tumor fragments obtained from 28 of the 40 CRC patients. The tumors were then dissociated into cell suspensions to be orthotopically injected into secondary mice. The tumors from 13 of the 28 patients progressed. Furthermore, metastases formed spontaneously in the liver and lungs from the tumor fragments obtained from 8 of these 13 patients. Moreover, employing a mathematical analysis, we showed that tumor cell clusters seeded these metastases significantly more often than did single tumor cells. Membrane E‐cadherin‐ and nuclear ZEB1‐positive tumor cells indicating the hybrid epithelial/mesenchymal state were also detected in primary tumors of various CRC patients, and in the corresponding patient‐derived xenografts (PDXs) and circulating tumor cell clusters in the bloodstreams of mice. In contrast, ZEB1 staining was barely detectable in the patient‐matched liver metastases presumably developing through mesenchymal‐to‐epithelial transition. Inhibition of E‐cadherin or ZEB1 expression by shRNA notably prevented the PDX‐derived tumor organoids from colonizing the liver, when injected intrasplenically into mice, indicating E‐cadherin and ZEB1 expressions to be required for their metastatic colonization. Taken together, these findings suggest that the epithelial/mesenchymal state mediates metastatic seeding of human CRC cell clusters into distant organs.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Establishment Of the Crc Pdx Mouse Models By Subcutaneous Anmentioning

confidence: 99%

Section: Establishment Of Experimental Tumor and Metastasis Models Ofmentioning

confidence: 99%

Section: Establishment Of Experimental Tumor and Metastasis Models Ofmentioning

confidence: 99%

See 3 more Smart Citations

Metastatic seeding of human colon cancer cell clusters expressing the hybrid epithelial/mesenchymal state

Mizukoshi

Okazawa

Haeno

et al. 2019

Intl Journal of Cancer

Self Cite

View full text Add to dashboard Cite

show abstract

Translational and Clinical Relevance of PDX‐Derived Organoid Models in Oncology Drug Discovery and Development

Kumari¹,

Li³

2022

Current Protocols

View full text Add to dashboard Cite

Patient‐derived cancer disease models conserve many key features of the original human cancers, potentially allowing higher predictive power than traditional cell line models. Accordingly, in vivo patient‐derived xenografts (PDX) are frequently utilized in preclinical and translational oncology studies as patient surrogates for population‐based screens (“mouse clinical trials”), for which large PDX biobanks have been generated over the last decade from various cancer types. In vitro patient‐derived organoids (PDO) have recently emerged as a disruptive technology, enabling early “patient in a dish” clinical trials. Like PDX, PDOs retain the histology/genomics of the original tumor and are highly predictive of the clinical response. Organoids derived from adult stem cells (ASC) in patient tissue can function as mini‐organs. They have greater advantages over other 3D in vitro systems, making them highly predictive, reliable, and consistent in vitro models. Large biobanks enable the adoption of organoids in early drug screening and patient selection. PDX biobanks, as a source of human material, have been used to create 3D in vitro screens, but with limitations. However, creating organoids from the ASCs residing in PDXs has been successfully used as a rapid and cost‐effective way to enable higher throughput in vitro screens and generate matched in vitro/in vivo model pairs that retain genomic, histopathological, and pharmacology profiles. This overview summarizes the generation of matched in vitro/in vivo models from patient material, the advantages over other systems, and the applications to drug discovery. © 2022 Wiley Periodicals LLC.

show abstract

A Tissue Engineering Approach to Metastatic Colon Cancer

et al. 2020

View full text Add to dashboard Cite

Colon cancer remains the third most common cause of cancer in the US, and the third most common cause of cancer death. Worldwide, colon cancer is the second most common cause of cancer and cancer deaths. At least 25% of patients still present with metastatic disease, and at least 25-30% will develop metastatic colon cancer in the course of their disease. While chemotherapy and surgery remain the mainstay of treatment, understanding the fundamental cellular niche and mechanical properties that result in metastases would facilitate both prevention and cure. Advances in biomaterials, novel 3D primary human cells, modelling using microfluidics and the ability to alter the physical environment, now offers a unique opportunity to develop and test impactful treatment. METASTATIC COLORECTAL CANCER: CLINICAL SIGNIFICANCE AND PATHOGENESIS Colorectal cancer (CRC) continues to grow in prevalence throughout the world and now accounts for nearly one in every ten cancers worldwide, constituting the second most common malignancy worldwide and the third most common in the United States (Bray et al., 2018) (Siegel et al., 2020). For colorectal cancer, stage at diagnosis is the most important predictor of survival with 5-year relative survival rates ranging from 90% in patients with local disease (stage I, II) to 14% for those diagnosed with distant (stage IV) disease (Bray et al., 2018; Siegel et al., 2020). Despite the plethora of research and the growing number of therapeutics, more than 700,000 deaths occur from CRC annually (Lozano et al., 2012).

show abstract

High-sensitivity Detection of Micrometastases Generated by GFP Lentivirus-transduced Organoids Cultured from a Patient-derived Colon Tumor

Cited by 7 publications

References 14 publications

Metastatic seeding of human colon cancer cell clusters expressing the hybrid epithelial/mesenchymal state

Metastatic seeding of human colon cancer cell clusters expressing the hybrid epithelial/mesenchymal state

Translational and Clinical Relevance of PDX‐Derived Organoid Models in Oncology Drug Discovery and Development

A Tissue Engineering Approach to Metastatic Colon Cancer

Contact Info

Product

Resources

About