NUT midline carcinoma as a primary lung tumor treated with anlotinib combined with palliative radiotherapy: a case report

Background NUT (nuclear protein in testis) midline carcinoma (NMC) is a rapidly progressive tumor arising from midline structures. Recent cases have reported that the poor prognosis with a median survival of 6.7 months and a 2 years overall survival of 19% due to limited treatment. Based on the effect of arotinib on inhibiting tumor growth and angiogenesis. We present one patient case treated with anlotinib and radiotherapy. Case presentation Here, we describe a 33-year old patient who complained of cough and chest pain and was diagnosed as a pulmonary NMC through CT scan, FISH and immunohistochemistry. In addition, we initially demonstrated that anlotinib combined with palliative radiotherapy could significantly prevent the tumor growth in a pulmonary NMC. Conclusion The report indicated that anlotinib combined with palliative radiotherapy could inhibit the tumor progression in a pulmonary NMC, which may provide a combined therapy to pulmonary NMC in the future.

NUT Midline Carcinoma Mimics Anaplastic Thyroid Carcinoma

Introduction/Objective NUT carcinoma (NC) is an extremely rare, aggressive subtype of squamous cell carcinoma defined by rearrangement of the NUTM1 (aka NUT) gene. NC most commonly arises within the thorax and head and neck and predominantly affects teens and young adults. For the first time, we described a rare case of NC mimics anaplastic thyroid carcinoma. Methods/Case Report A 27-year female with no significant past medical history presented with neck pain and swelling for a few months and was noted to have a “thyroid” mass. An ultrasound guided fine needle aspiration of the mass showed a hypercellular specimen composed of highly atypical epithelial cells with irregular nuclear membrane and prominent nucleolus, high nuclear to cytoplasmic ration, dense cytoplasm, lying singly or in flat sheet mixed with acute inflammatory exudate (Fig. A, B). No thyroid follicular cells or colloid are present. It was signed out as suspicious for squamous cell carcinoma since there was no cell block for further workup. A following excisional biopsy demonstrated similar atypical epithelium detached or embedded in skeletal muscle and soft tissue with acute, chronic inflammation and necrosis (Fig C). By immunohistochemical stains, the atypical cells are positive for p40 (Fig D) and p53. The Ki-67 proliferation index is ranging from 40 % to 70 %. Scattered tumor cells are weakly positive for PAX-8 and TTF-1. These findings raise the possibility that the squamous cell carcinoma might be a component of anaplastic thyroid carcinoma. However, molecular studies revealed a NSD3-NUTM1 fusion, which is characteristically identified in a subtype of squamous cell carcinoma know as NUT “midline” carcinoma. Results (if a Case Study enter NA) NA Conclusion Diagnosis of NC can be established by positive NUT nuclear immunohistochemical staining. Recognizing the typical morphology and keeping low threshold for the NUT immunohistochemical staining will increase the diagnostic efficacy.

SETD2 Deficiency and miR-21: Potent Therapeutic Targets in NUT Midline Carcinoma

BackgroundNuclear protein in testis (NUT) midline carcinoma (NMC) is a rare and highly aggressive tumor with the bromodomain containing 4 protein (BRD4)-NUT (NUTM1) gene fusion. Few targeted therapies are available for NMC, and novel therapeutic targets are required. The objectives of our study was to determine gene mutations and microRNA (miRNA) expression levels in NMC and to identify novel therapeutic targets for NMC. Methods Next-generation sequencing (NGS) was used to identify mutations in the NMC cell lines and the tumor sample from a NMC patient; we determined the BRD4-NUT fusion gene in the patient using a digital PCR assay. Trimethylation of lysine 36 on histone H3 expression (H3K36me3) was studied using western blotting. The efficacy of the WEE1 G2 checkpoint kinase (WEE1) inhibitor, AZD1775, was evaluated using the MTS assay. RNA sequencing was performed to determine miRNA expression levels in the NMC cell lines. TaqMan miRNA assays were used to analyze miR-21 expression. The efficacy of miR-21 inhibitor was evaluated using the MTS assay. Results A novel SETD2 mutation (p.Ser2382fs) was identified in the NMC cell lines and the patient tumor. H3K36me3 was depleted in the NMC cells, which was indicative of functional SETD2 loss. The NMC cells were sensitive to AZD1775, an inhibitor for SETD2-deficient cancer. miRNA analysis revealed that miR-21 expression was increased in the NMC cells resistant to the traditional BET-targeted therapy. The miR-21 inhibitor suppressed the proliferation of the NMC cells. ConclusionsSETD2 deficiency and miR-21 may serve as novel therapeutic targets for the treatment of NMC.Trial registrationIn this study, we analyzed the gene alterations in human tumor samples. This study was registered in the UMIN clinical trial registered system (UMIN000043147, January 27, 2021).

Case Report and Literature Review: Primary Pulmonary NUT-Midline Carcinoma

Nuclear protein of the testis (NUT) carcinoma is a very rare and aggressive carcinoma characterized by chromosomal rearrangement. NUT-midline carcinoma (NMC) can occur anywhere in the body, but most of the tumors are found in the midline anatomic structure or mediastinum. Pulmonary-originated NMC is extremely rare and often difficult to be distinguished from other poorly differentiated tumors, making the diagnosis awfully challenged in clinical practice. There are less than 100 cases of NUT carcinoma reported so far. In this study, the diagnosis and molecular mechanisms of reported NUT carcinoma cases were reviewed. Furthermore, a case of primary pulmonary NUT-midline carcinoma and its pathological features was reported. The process of pathological identification and genomic analysis for establishing the diagnosis was discussed. We found that NUT carcinoma could be identified by combining CT, H&E staining, immunohistochemistry (IHC), and molecular tests. The development of NUT carcinoma might be associated with mutation of MYC, p63, and MED24 genes and the Wnt, MAPK, and PI3K signaling pathways. Our study provided a detailed molecular mechanistic review on NMC and established a procedure to identify pulmonary NMC.

SETD2 and miR-21 as therapeutic targets for NUT midline carcinoma

Background:Nuclear protein in testis (NUT) midline carcinoma (NMC) is a rare and highly aggressive tumor with the bromodomain containing 4 (BRD4)-NUT (NUTM1) gene fusion. BRD4 is a member of the bromodomain and extra-terminal domain (BET) family of proteins, and BET inhibitors have been investigated in NMC clinical trials. However, few targeted therapies are available for NMC, and novel therapeutic targets remain to be determined. We determined the role of two epigenetic regulators as possible therapeutic targets for NMC. Methods:We performed next-generation sequencing (NGS) in NMC cell lines (HCC2429 and Ty82). H3K36me3 expression was studied using western blotting. The efficacy of AZD1775, a WEE1 inhibitor, was evaluated using the MTS and γH2AX assays. We established an NMC cell line that was resistant to BET inhibitors. The sensitivity of the cells resistant to AZD1775 was analyzed using the MTS assay. RNA sequencing was performed to determine miRNA expression levels. TaqMan miRNA assays were used to analyze miR-21 expression. The efficacy of the miR-21 inhibitor was evaluated using the MTS assay. We established a digital PCR (dPCR) assay to detect NUT gene rearrangements to identify patients with NMC. Using NGS, a patient with NMC was identified with the SETD2 mutation.Results:SETD2 mutation (p.Ser2382fs) was determined in NMC cells, in which H3K36me3 expression was depleted, indicating SETD2 loss. NMC cells were sensitive to the WEE1 inhibitor, AZD1775 in cancer cells with SETD2 deficiency. γH2AX expression was increased in NMC cells treated with AZD1775. The efficacy of the combination of AZD1775 and JQ-1 was additive. We established NMC cells that were resistant to BET inhibitors. The resistant cells were also sensitive to AZD1775. miRNA analysis revealed increased miR-21 expression in BET-inhibitor-resistant NMC cells. MiR-21 regulated the growth of NMCs. The miR-21 inhibitor suppressed the growth of BET-inhibitor-resistant cells. Thirty-two clinical samples were analyzed and NMC was identified using digital PCR assays. Tumors with the SETD2 mutation were analyzed using NGS.Conclusions:We report for the first time SET domain-containing protein 2 (SETD2) deficiency and miR-21 as novel therapeutic targets for NMC. SETD2 loss and miR-21 may be therapeutic targets for NMC.Trial registrationIn this study, we analyzed the gene alterations in human tumor samples. This study was registered in the UMIN clinical trial registered system (UMIN000043147, January 27, 2021).