bims-myxlip 2021-08-01 papers

Issue of 2021‒08‒01
five papers selected by
Laura Mannarino
Humanitas Research

Hum Cell. 2021 Jul 24.

Establishment and characterization of novel patient-derived cell lines from giant cell tumor of bone.

Yuki Yoshimatsu, Rei Noguchi, Ryuto Tsuchiya, Takuya Ono, Yooksil Sin, Sei Akane, Jun Sugaya, Tomoaki Mori, Suguru Fukushima, Akihiko Yoshida, Akira Kawai, Tadashi Kondo.

Giant cell tumor of bone (GCTB) is a locally aggressive and rarely metastasizing tumor. GCTB is characterized by the presence of unique giant cells and a recurrent mutation in the histone tail of the histone variant H3.3, which is encoded by H3F3A on chromosome 1. GCTB accounts for ~ 5% of primary bone tumors. Although GCTB exhibits an indolent course, it has the potential to develop aggressive behaviors associated with local recurrence and distant metastasis. Currently, complete surgical resection is the only curative treatment, and novel therapeutic strategies are required. Patient-derived cancer cell lines are critical tools for basic and pre-clinical research. However, only a few GCTB cell lines have been reported, and none of them are available from public cell banks. Therefore, we aimed to establish novel GCTB cell lines in the present study. Using curetted tumor tissues of GCTB, we established two cell lines and named them NCC-GCTB2-C1 and NCC-GCTB3-C1. These cells harbored a typical mutation in histones and exhibited slow but constant growth, formed spheroids, and had invasive capabilities. We demonstrated the utility of these cell lines for high-throughput drug screening using 214 anticancer agents. We concluded that NCC-GCTB2-C1 and NCC-GCTB3-C1 cell lines were useful for the in vitro study of GCTB.

Keywords: Cell lines; Giant cell tumor of bone; High-throughput screening; Patient-derived cancer model; Spheroids

DOI: https://doi.org/10.1007/s13577-021-00579-z

Cell Death Dis. 2021 Jul 29. 12(8): 749

PIK3R3, part of the regulatory domain of PI3K, is upregulated in sarcoma stem-like cells and promotes invasion, migration, and chemotherapy resistance.

Changhwan Yoon, Jun Lu, Sandra W Ryeom, M Celeste Simon, Sam S Yoon.

To identify drivers of sarcoma cancer stem-like cells (CSCs), we compared gene expression using RNA sequencing between HT1080 fibrosarcoma and SK-LMS-1 leiomyosarcoma spheroids (which are enriched for CSCs) compared with the parent populations. The most overexpressed survival signaling-related gene in spheroids was phosphoinositide-3-kinase regulatory subunit 3 (PIK3R3), a regulatory subunit of PI3K, which functions in tumorigenesis and metastasis. In a human sarcoma microarray, PIK3R3 was also overexpressed by 4.1-fold compared with normal tissues. PIK3R3 inhibition using shRNA in the HT1080, SK-LMS-1, and DDLS8817 dedifferentiated liposarcoma in spheroids and in CD133+ cells (a CSC marker) reduced expression of CD133 and the stem cell factor Nanog and blocked spheroid formation by 61-71%. Mechanistic studies showed that in spheroid cells, PIK3R3 activated AKT and ERK signaling. Inhibition of PIK3R3, AKT, or ERK using shRNA or inhibitors decreased expression of Nanog, spheroid formation by 68-73%, and anchorage-independent growth by 76-91%. PIK3R3 or ERK1/2 inhibition similarly blocked sarcoma spheroid cell migration, invasion, secretion of MMP-2, xenograft invasion into adjacent normal tissue, and chemotherapy resistance. Together, these results show that signaling through the PIK3R3/ERK/Nanog axis promotes sarcoma CSC phenotypes such as migration, invasion, and chemotherapy resistance, and identify PIK3R3 as a potential therapeutic target in sarcoma.

DOI: https://doi.org/10.1038/s41419-021-04036-5

Pathol Res Pract. 2021 Jul 21. pii: S0344-0338(21)00216-8. [Epub ahead of print]225 153555

The utility of fluorescence in situ hybridization (FISH) in determining DNA damage-inducible transcript 3 (DDIT3) amplification in dedifferentiated liposarcomas - an important diagnostic pitfall.

Olga Kuczkiewicz-Siemion, Piotr Wiśniewski, Agnieszka Dansonka-Mieszkowska, Magdalena Grabowska-Kierył, Katarzyna Olszewska, Tomasz Goryń, Monika Prochorec-Sobieszek, Piotr Rutkowski, Anna Szumera-Ciećkiewicz.

BACKGROUND AND OBJECTIVE: Dedifferentiated liposarcoma (DDLPS) is characterized by non-lipogenic sarcoma fields coexisting with adipocyte-rich well-differentiated areas. Amplification of the 12q13-15 region includes the MDM2 and DDIT3 genes. MDM2 amplification is considered a genetic hallmark of DDLPS, while DDIT3 is typically rearranged in myxoid liposarcoma. Recent studies showed that DDIT3 amplification is associated with myxoid liposarcoma-like (LPS-like) morphology in DDLPS. Our study aimed to evaluate the status of MDM2 and DDIT3 by FISH in DDLPS and correlate it with MLPS-like features.MATERIAL AND METHODS: Six patients with MLPS-like morphology DDLPS were investigated pathologically, immunohistochemically, and genetically. The control groups of patients with classical DDLPS morphology and well-differentiated liposarcoma (WDLPS) were established and molecularly assessed as well. Fluorescence in situ hybridization (FISH) used in routine diagnostics was performed to determine the status of MDM2 and DDIT3 genes.
RESULTS: The patient's mean age was 64 (range from 43 to 85 years) with a 5:4 male to female ratio. Tumors were localized retroperitoneally (15) and extra-retroperitoneally (3). All cases demonstrated amplification of the 12q15 region containing MDM2 gene and co-amplification of the 5' DDIT3 FISH Probe representing DDIT3 telomeric tag. However, we did not find the relation of myxoid LPS-like morphology with DDIT3 amplification as previously reported.
CONCLUSIONS: The biopsy material from DDLPS with myxoid areas can be misclassified as myxoid liposarcoma. Indeed, according to the histological image, DDIT3 status may be evaluated first. In these cases, we show that the DDIT3 telomeric tag amplification assessed by FISH, is a common, nonspecific feature, which is also found in classical DDLPS and WDLPS. Therefore, we believe that co-amplification of DDIT3 and MDM2 may be considered a spectrum of the 12q13-15 region amplification due to the specification of FISH methodology.

Keywords: DDIT3 amplification; Dedifferentiated liposarcoma; Fluorescence in situ hybridization; MDM2 amplification; Myxoid liposarcoma-like features

DOI: https://doi.org/10.1016/j.prp.2021.153555

Mol Cancer Ther. 2021 Jul 26. pii: molcanther.MCT-20-0489-A.2020. [Epub ahead of print]

Pharmaceutical interference of the EWS-FLI1-driven transcriptome by co-targeting H3K27ac and RNA polymerase activity in Ewing Sarcoma.

Daniel A R Heisey, Sheeba Jacob, Timonthy L Lochmann, Richard Kurupi, Maninderjit S Ghotra, Marissa L Calbert, Mayuri Shende, Yuki Kato Maves, Jennifer E Koblinski, Mikhail G Dozmorov, Sosipatros A Boikos, Cyril H Benes, Anthony C Faber.

The EWSR1-FLI1 t(11;22)(q24;q12) translocation is the hallmark genomic alteration of Ewing Sarcoma (ES), a malignancy of the bone and surrounding tissue, predominantly affecting children and adolescents. Although significant progress has been made for the treatment of localized disease, patients with metastasis or who relapse after chemotherapy have less than a 30% five-year survival rate. EWS-FLI1 is currently not pharmaceutically druggable, driving the need for more effective targeted therapies. Treatment with the H3K27 demethylase inhibitor, GSK-J4, leads to an increase in H3K27me and a decrease in H3K27ac, a significant event in ES because H3K27ac associates strongly with EWS-FLI1 binding at enhancers and promoters and subsequent activity of EWS-FLI1 target genes. We were able to identify targets of EWS-FLI1 tumorigenesis directly inhibited by GSK-J4. GSK-J4 disruption of EWS-FLI1-driven transcription was toxic to ES cells and slowed tumor growth in PDXs of ES. Responses were markedly exasperated by co-treatment with a disruptor of RNA polymerase II activity, the CDK7 inhibitor THZ1. This combination together suppressed EWS-FLI1 target genes and viability of ex vivo PDX ES cells in a synergistic manner. In a PDX model of ES, the combination shrank tumors. We present a new therapeutic strategy to treat ES by decreasing H3K27ac at EWS-FLI1-driven transcripts, exasperated by blocking phosphorylation of the C-terminal domain of RNA polymerase II to further hinder the EWS-FLI1-driven transcriptome.

DOI: https://doi.org/10.1158/1535-7163.MCT-20-0489