Three-dimensional Coculture Provides an Improved in Vitro Model for Papillary Renal Cell Carcinoma

2021 ◽  
Author(s):  
Kylee A Rosette ◽  
Stephen M Lander ◽  
Calvin VanOpstall ◽  
Brendan David Looyenga
Keyword(s):  
Renal Cell Carcinoma ◽  
Cell Carcinoma ◽  
Tumor Cells ◽  
Renal Cell ◽  
Three Dimensional ◽  
Human Tumor ◽  
Papillary Renal Cell Carcinoma ◽  
Specific Pattern ◽  
Stromal Fibroblasts

Papillary renal cell carcinoma (pRCC) represents the second most common kidney cancer and can be distinguished from other types based upon its unique histologic architecture and specific pattern of genomic alterations. Sporadic Type1 pRCC is almost universally driven by focal or chromosomal amplification of the receptor tyrosine kinase MET, though the specific mode of its activation is unclear. While the MET receptors found in human tumor specimens appear highly active, those found on the surface of in vitro cultured tumor cells are only weakly activated in the absence of exogenous hepatocyte growth factor (HGF) ligand. Furthermore, pRCC cells cultured in standard two-dimensional conditions with serum fail to respond functionally to MET knockdown or the selective MET inhibitor capmatinib despite clear evidence of kinase inhibition at the molecular level. To better model pRCC in vitro, we developed a three-dimensional (3D) coculture system in which renal tumor cells are layered on top of primary fibroblasts in a fashion that mimics the papillary architecture of human tumors. In this 3D spheroid model, the tumor cells survive and proliferate in the absence of serum due to trophic support of HGF-producing fibroblasts. Unlike tumor cells grown in monoculture, the proliferation of cocultured tumor cells is sensitive to capmatinib and parallels inhibition of MET kinase activity. These findings demonstrate the importance of stromal fibroblasts in pRCC and indicate that accurate in vitro representation of this disease requires the presence of both tumor and fibroblast cells in a structured coculture model.

scholarly journals Efficacy of povidone-iodine against accidental tumor incision during nephron-sparing surgery: experimental study in patients with renal cell carcinoma

2019 ◽  
Vol 47 (10) ◽  
pp. 4993-5002
Author(s):  
Gang Li ◽  
Chao Zhi ◽  
Dongsheng Zhu ◽  
Zihao Liu ◽  
Yuanjie Niu
Keyword(s):  
Renal Cell Carcinoma ◽  
Local Recurrence ◽  
Cell Carcinoma ◽  
Tumor Cells ◽  
Renal Cell ◽  
Povidone Iodine ◽  
Nephron Sparing Surgery ◽  
In Vitro Experiments ◽  
Nephron Sparing

Purpose Accidental tumor incision (ATI) can occur during nephron-sparing surgery (NSS) and correlates with recurrence and metastasis. This study investigated risk factors of intraoperative ATI in renal cell carcinoma (RCC) patients after NSS and the efficacy of povidone-iodine for ATI. Methods A retrospective analysis was performed on 150 consecutive stage I (pT1N0M0) RCC patients who underwent NSS at The Second Hospital of Tianjin Medical University between May 2010 and October 2015 for the causes of ATI. Furthermore, in vitro experiments investigated whether tumor cells remained on the surface of scissors and the effect of treatment with povidone-iodine on the number of remaining 786-O cells. Results Among the 150 cases, 15 showed ATI, of which three suffered local recurrence during a median follow-up of 56 months. Pseudocapsules, satellite nodules, and renal cystic tumors were observed in ATI cases. In vitro experiments showed that tumor cells remained on the surface of scissors after ATI during NSS and that 0.5% povidone-iodine effectively killed tumor cells in 30 minutes. Conclusions The probability of ATI is high in patients with complex-type RCC during NSS. ATI potentially increases the chance of metastasis and local recurrence, and 0.5% povidone-iodine kills tumor cells more effectively than distilled water.

FGFR2 expression to predict survival outcome in patients with metastatic papillary renal cell carcinoma.

2016 ◽  
Vol 34 (2_suppl) ◽  
pp. 506-506 ◽  
Author(s):  
Ilya Tsimafeyeu ◽  
Alexandra Naumova ◽  
Evgenia Stepanova ◽  
Alfia Khasanova ◽  
Ilya Varlamov ◽  
...  
Keyword(s):  
Renal Cell Carcinoma ◽  
Cell Carcinoma ◽  
Tumor Cells ◽  
Renal Cell ◽  
Objective Response ◽  
Progression Free Survival ◽  
Papillary Renal Cell Carcinoma ◽  
Survival Outcome ◽  
Primary Tumors ◽  
Metastatic Sites

506 Background: In our previous study we showed that fibroblast growth factor receptor 2 (FGFR2) mutations are rare across papillary types of renal cell carcinoma (pRCC). The aim of the present study is to test FGFR2 expression for association with survival outcome in the largest patient cohort to date. Methods: Formalin-fixed, paraffin-embedded specimens of removed primary tumors from 214 untreated metastatic pRCC patients were evaluated by immunohistochemistry with FGFR2 antibody (Santa Cruz Biotechnology). Expression was quantified by consensus of two independent observers using a four-value intensity score (0, 1+, 2+, and 3+) and the percentage (0-100%) of the extent of reactivity. Expression was scored according to the percentage of positive cells present among all tumor cells in the section. The cytoplasmic and nuclear expression score was obtained by multiplying the intensity and reactivity extension values (range, 0-300). FGFR2 expression was tested for associations with progression-free survival (PFS), overall survival (OS) and best objective response. Results: Expression of FGFR2 was observed in 23% (49/214) of primary pRCC, mostly in cytoplasm of tumor cells. 2 of 214 (1%) patients had nuclear FGFR2 expression. Intensity was 3+ in all cases. Expression of FGFR2 was significant lower in the normal tissue of kidney (1%, P=0.001). FGFR2 expression was strongly associated with a number of metastatic sites (2 and more metastatic sites vs. 0-1), type 2 of pRCC, lower nucleolar grade (P<0.001). FGFR2-positive patients had significantly shorter OS and PFS in first-line therapy (P<0.05; Table). On multivariate analysis, FGFR2 expression, MSKCC risk group, and type of pRCC were found to be independent predictors of survival. Conclusions: In this study, we described immunohistochemical expression of FGFR2 in a large series of pRCC specimens. FGFR2 expression was found to be prognostic factor for survival in patients with metastatic pRCC. Clinical trial information: rosoncoweb2011. [Table: see text]

scholarly journals Prognostic and Therapeutic Potential of The OIP5 Network in Papillary Renal Cell Carcinoma

2021 ◽  
Author(s):  
Mathilda Chow ◽  
Yan Gu ◽  
Lizhi He ◽  
Xiaozeng Lin ◽  
Ying Dong ◽  
...  
Keyword(s):  
Renal Cell Carcinoma ◽  
Cell Carcinoma ◽  
Renal Cell ◽  
Therapeutic Potential ◽  
Synthetic Lethality ◽  
Papillary Renal Cell Carcinoma ◽  
Mrna Levels ◽  
Rna Seq

Abstract Background: Papillary renal cell carcinoma (pRCC) is an aggressive but minor type of RCC compared to the main RCC type, clear cell RCC. The current understanding and management of pRCC remain poor. OIP5 possesses oncogenic functions; its contributions to pRCC remain unknown.Methods: OIP5 expression in pRCC at both the protein and mRNA levels was determined using tissue microarray and TCGA dataset. OIP5 was ectopically expressed in metastatic ACHN pRCC cells; xenografts were performed with gene expression profiled by RNA-seq. Differentially expressed genes (DEGs) were analyzed for prognostic potential and impact on pRCC. The effect of PLK1, an OIP5-related DEG, on pRCC tumor growth in vivo was examined.Results: OIP5 expression is upregulated in pRCC. The upregulation associates with pRCC adverse features (T1P<T2P<CIMP, Stage1+2<Stage 3<Stage 4, and N0<N1) and effectively stratifies the fatality risk. OIP5 promotes ACHN pRCC cell proliferation and xenograft formation. RNA-seq reveals network alterations related to immune regulation, metabolism, and hypoxia in ACHN OIP5 tumors compared to empty vector tumors. A set of DEGs was derived from ACHN OIP5 xenografts and primary pRCCs (n=282) contingent to OIP5 upregulation; both DEG sets share 66 overlap genes. Overlap66 effectively predicts overall survival (p<2e-16) and relapse (p<2e-16) possibilities. The prediction is associated with a good out-of-sample performance, supporting its clinical applications. High-risk tumors stratified by Overlap66 risk score possess an immune suppressive environment, evident by elevations in Treg cells and PD1 expression in CD8 T cells. Upregulation of PLK1 occurs in both xenografts and primary pRCC tumors with OIP5 elevations. PLK1 displays a synthetic lethality relationship with OIP5; PLK1 inhibitor BI2356 causes G2/M arrest in ACHN OIP5 cells in vitro and significantly inhibits the growth of xenografts formed by ACHN OIP5 cells in vivo. Conclusions: Our research reveals that OIP5 and its network possess robust prognostic and therapeutic potentials; the prognostic value of Overlap66 and the therapeutic potential of PLK1 inhibitors may pave the way for developing personalized medicine for pRCC management.

scholarly journals Cabozantinib Reverses Renal Cell Carcinoma–mediated Osteoblast Inhibition in Three-dimensional Coculture In Vitro and Reduces Bone Osteolysis In Vivo

2020 ◽  
Vol 19 (6) ◽  
pp. 1266-1278
Author(s):  
Tianhong Pan ◽  
Mariane Martinez ◽  
Kelsea M. Hubka ◽  
Jian H. Song ◽  
Song-Chang Lin ◽  
...  
Keyword(s):  
Renal Cell Carcinoma ◽  
Cell Carcinoma ◽  
Renal Cell ◽  
Three Dimensional

scholarly journals Adult Nephroblastoma with Predominant Epithelial Component: A Differential Diagnostic Candidate of Papillary Renal Cell Carcinoma and Metanephric Adenoma—Report of Three Cases

2013 ◽  
Vol 2013 ◽  
pp. 1-5
Author(s):  
Shiho Watanabe ◽  
Hiroshi Naganuma ◽  
Michio Shimizu ◽  
Satoshi Ota ◽  
Shin-ichi Murata ◽  
...  
Keyword(s):  
Renal Cell Carcinoma ◽  
Cell Carcinoma ◽  
Tumor Cells ◽  
Renal Cell ◽  
Benign Tumor ◽  
Renal Tumor ◽  
Papillary Renal Cell Carcinoma ◽  
Metanephric Adenoma ◽  
Old Females ◽  
Epithelial Element

Although nephroblastoma is the commonest renal tumor of childhood, it is rare in adults. In cases of predominantly epithelial type occurring in adulthood, it might be difficult to distinguish it from papillary renal cell carcinoma and metanephric adenoma. Here, we report three cases of adult epithelial nephroblastoma in 24-, 76-, and 21-year-old females. Histologically, the tumors were composed of papillotubular architectures of small and uniform tumor cells with high nucleocytoplasmic ratio without blastemal element. Immunohistochemically, the tumor cells were positive for WT-1 and CD57 but negative for AMACR, which was helpful to exclude the possibility of papillary renal cell carcinoma. Metanephric adenoma is a benign tumor, which can be distinguished by the observation of the cellular atypism and growth pattern. However, nephroblastoma with predominant epithelial element mimics the malignant counterpart of metanephric adenoma, that is, “metanephric adenocarcinoma.”

Type 1 papillary renal cell carcinoma in a patient with schwannomatosis: Mosaic versus loss of SMARCB1 expression in respectively schwannoma and renal tumor cells

2016 ◽  
Vol 55 (4) ◽  
pp. 350-354 ◽  
Author(s):  
Theo J.M. Hulsebos ◽  
Susan Kenter ◽  
Frank Baas ◽  
Eline A. Nannenberg ◽  
Fonnet E. Bleeker ◽  
...  
Keyword(s):  
Renal Cell Carcinoma ◽  
Cell Carcinoma ◽  
Tumor Cells ◽  
Renal Cell ◽  
Renal Tumor ◽  
Papillary Renal Cell Carcinoma

scholarly journals A mutated HLA-A2 molecule recognized by autologous cytotoxic T lymphocytes on a human renal cell carcinoma.

1996 ◽  
Vol 183 (6) ◽  
pp. 2501-2508 ◽  
Author(s):  
D Brändle ◽  
F Brasseur ◽  
P Weynants ◽  
T Boon ◽  
B Van den Eynde
Keyword(s):  
Renal Cell Carcinoma ◽  
T Lymphocytes ◽  
Cell Carcinoma ◽  
Tumor Cells ◽  
Cytotoxic T Lymphocytes ◽  
Renal Cell ◽  
Human Tumor ◽  
Alpha Helix ◽  
Human Renal Cell Carcinoma ◽  
Hla Class I Molecules

Many human tumor cells have been shown to express antigens that are recognized by autologous cytotoxic T lymphocytes (CTL) and the molecular nature of a number of melanoma antigens has been defined recently. Here we describe the characterization of an antigen recognized on a renal cell carcinoma by autologous CTL clones. This antigen is encoded by the HLA-A2 gene present in the tumor cells. The sequence of this gene differs from the HLA-A2 sequence found in autologous peripheral blood lymphocytes by a point mutation that results in an arginine to isoleucine exchange at residue 170, which is located on the alpha-helix of the alpha 2 domain. Transfection experiments with the normal and mutated HLA-A2 cDNA demonstrated that this amino acid replacement was responsible for the recognition of the HLA-A2 molecule expressed on the tumor cells. The mutant HLA-A2 gene was also detected in the original tumor tissue from the patient, excluding the possibility that the mutation had appeared in vitro. Thus, HLA class I molecules carrying a tumor-specific mutation can be involved in the recognition of tumor cells by autologous CTL.

In Vitro-Expanded NK Cells Have Increased TRAIL and NKG2D Expression and Enhanced TRAIL-Mediated Tumor Cytotoxicity Compared to Non-Expanded NK Cells.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 2744-2744 ◽  
Author(s):  
Maria Berg ◽  
Andreas Lundqvist ◽  
Yong Fan ◽  
J. Philip McCoy ◽  
Hisayuki Yokoyama ◽  
...  
Keyword(s):  
Renal Cell Carcinoma ◽  
Cell Carcinoma ◽  
Nk Cells ◽  
Tumor Cells ◽  
Renal Cell ◽  
Nk Cell ◽  
Surface Expression ◽  
Renal Cell Carcinoma Cell ◽  
Tumor Cytotoxicity

Abstract The activation of NK cell inhibitory receptors may limit the antitumor efficacy of adoptive autologous and allogeneic NK cell infusions. Recently, we found that exposing malignant cells to the proteosome inhibitor bortezomib upregulated surface expression of death receptors for TRAIL, resulting in significant enhancement of autologous NK cell tumor cytotoxicity in vitro and in vivo. Here we show that NK cells expanded in vitro in the presence of IL-2 and EBV-LCL feeder cells upregulate surface expression of TRAIL which significantly augments bortezomib-induced tumor sensitization to NK cell killing. CD56+/CD3– NK cells were isolated from normal donors by immuno-magnetic bead selection and were co-cultured with irradiated EBV-LCL feeder cells in X-VIVO 20, 10% human AB serum, and 500 IU/ml hrIL-2 for up to 21 days. Depending on culture conditions, a 300 to 10,000 fold increase in NK cell numbers was achieved. Non-expanded and expanded NK cells were analyzed by flow cytometry for the expression of CD56, CD16, TRAIL, FasL, NKG2D, LFA-1, perforin, and granzymes A and B at baseline and ≥ 10 days following in vitro expansion. Chromium release assays were performed to assess fresh vs. expanded NK cell cytotoxicity of renal cell carcinoma (RCC) tumor targets treated with 10 nM bortezomib for 18 hr vs. untreated RCC controls. Freshly-isolated NK cells did not express TRAIL or FasL; in contrast NKG2D, LFA-1, perforin and granzymes A and B were constitutively expressed in fresh NK cells. After expansion, there was a dramatic increase in surface expression of TRAIL and NKG2D; on fresh vs. expanded NK cells from 3 different donors, TRAIL expression increased from 0% to 80.8±15.4% (mean fluorescence intensity [MFI] of TRAIL increased from 6.0±5.1 to 37.9±3.2). The MFI of NKG2D surface expression also increased following NK cell expansion (432.0±70.9 from 48.3±16.3). Expression of LFA-1 and perforin did not change, although there was a small increase in surface and intracellular expression of FasL and granzymes A and B respectively. At a 1:1 effector to target ratio, fresh NK cells lysed 3.4± 2.1% and 5.0± 2.7% of untreated and bortezomib-treated RCC tumor cells respectively. In contrast, there was a dramatic increase in bortezomib-treated tumor susceptibility to killing by expanded NK cells; NK cells expanded for 12-18 days killed 27.6± 9.3% and 55.8± 8.3% of untreated vs. bortezomib-treated RCC tumor cells respectively. Conclusion: In vitro-expanded NK cells are phenotypically and functionally different from non-expanded NK cells. Expanded cells have increased NKG2D and TRAIL expression and greatly enhanced TRAIL-mediated tumor cytotoxicity compared to non-expanded NK cells. Based on these findings, a phase I trial investigating the safety and anti-tumor effects of escalating doses of adoptively-infused ex vivo-expanded autologous NK cells following bortezomib treatment in patients with advanced metastatic tumors and hematological malignancies has recently been initiated. Figure: Freshly isolated and expanded Nk cell lysis of renal cell carcinoma cell line with and without treatment of tumor cells with bortezomib Figure:. Freshly isolated and expanded Nk cell lysis of renal cell carcinoma cell line with and without treatment of tumor cells with bortezomib

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