西尾 永司

Abstract Approximately 660,000 women are diagnosed with cervical cancer annually. Current screening options such as cytology or human papillomavirus testing have limitations, creating a need to identify more effective ancillary biomarkers for triage. Here, we evaluated whether metabolomic analysis of cervical mucus metabolism could be used to identify biomarkers of cervical intraepithelial neoplasia (CIN) and cervical cancer. The case–control group consisted of 181 CIN, 69 squamous cell carcinoma (SCC) patients, and 48 healthy controls in the primary cohort. We undertook metabolomic analyses using ultra‐HPLC–tandem mass spectrometry. Univariate and multivariate analyses were carried out to profile metabolite characteristics, and receiver operating characteristic (ROC) analysis identified biomarker candidates. Five metabolites conferred the highest discriminatory power for SCC: oxidized glutathione (GSSG) (area under the ROC curve, 0.924; 95% confidence interval, 0.877–0.971), malic acid (0.914, 0.859–0.968), kynurenine (0.884, 0.823–0.945), GSSG/glutathione (GSH) (0.936, 0.892–0.979), and kynurenine/tryptophan (0.909, 0.856–0.961). Malic acid was the best marker for detection of CIN2 or worse (0.858, 0.793–0.922) and was a clinically useful metabolite. We confirmed the reproducibility of the results by validation cohort. Additionally, metabolomic analyses revealed eight pathways strongly associated with cervical neoplasia. Of these, only the tricarboxylic acid cycle was strongly associated with all CINs and cancer, indicating active energy production. Aberrant arginine metabolism by decreasing arginine and increasing citrulline might reduce tumor immunity. Changes in cysteine‐methionine and GSH pathways might drive the initiation and progression of cervical cancer. These results suggest that metabolic analysis can identify ancillary biomarkers and could improve our understanding of the pathophysiological mechanisms underlying cervical neoplasia.

Abstract Currently, human papillomavirus tests and cytology are used to screen for cervical cancer. However, more accurate ancillary screening tests are needed. MicroRNAs (miRNAs) and cytokines are promising biomarkers that are aberrantly expressed in cervical cancer. Therefore, the potential of developing new screening markers based on the levels of miRNAs and cytokines in serum and local mucus samples from the same patients with cervical neoplasia was investigated. miRNA screening was performed by microarray and measurement using real‐time reverse‐transcriptase PCR. Cytokine were measured using multiplex bead assay, and changes in expressions were analyzed based on disease severity. As lesions progressed, miR‐20b‐5p, −155‐5p, −144‐3p, −451a, and −126‐3p expression levels were increased in mucus, and miR‐16‐5p, −223‐3p, and ‐451a expression levels were decreased in serum. Regarding cytokines, IL‐6, IL‐8, monocyte chemoattractant protein‐1, Eotaxin, interferon‐γ, and RANTES were increased, whereas granulocyte–colony‐stimulating factor (G‐CSF) was significantly decreased in mucus. miRNAs and cytokines in serum did not have high diagnostic accuracy. However, a combination of miR‐20b‐5p, ‐451a, ‐126‐3p, Eotaxin, as well as G‐CSF in mucus samples, had high diagnostic accuracy with an area under the receiver operating characteristic curve of 0.989 (0.979–0.999). Our results suggest that using mucus for this ancillary test is more beneficial than serum.

INTRODUCTION: The da Vinci SP surgical system is a surgical platform capable of implementing robotic-assisted surgery through a single port and was first introduced in Japan at our hospital. In this paper, we describe our experience of the initial introduction of the da Vinci SP surgical system and its surgical outcomes. This is the first report on the surgical outcomes of using da Vinci SP, and its comparison with the conventional system in Japan. METHODS: After developing an application for a highly difficult new medical technology in-house, we compared the surgical outcomes (median values) of 15 patients who had undergone total hysterectomy at our hospital using the da Vinci SP (1-port) system (SP group) for uterine myoma after March 2023 and of 154 patients who underwent total hysterectomy using the conventional da Vinci Xi (four ports) system (Xi group) for uteri weighing <500 g. RESULTS: The results of the comparison of the characteristics between 15 patients in the SP group and 154 patients in the Xi group were as follows: uterus weight (g): 230 (90-500) versus 222 (55-496) (p = .35); surgical time (minutes): 199 (171-251) versus 198 (88-387) (p = .63); intraoperative blood loss (mL): 13 (5-82) versus 20 (2-384) (p = .17); and rate of surgical complication (%): 0.0 versus 1.3 (p = .66). The data indicated a comparable weight of the resected uterus, surgical time, intraoperative blood loss, and rate of surgical complications between the two groups. CONCLUSION: Robotic-assisted total hysterectomy using the da Vinci SP surgical system allowed clinicians to safely perform surgeries according to the conventional systems.

BACKGROUND/AIM: The frequency rate of injection site reactions (ISR) due to fosaprepitant meglumine (Fos APR) has been shown to vary depending on the types of combined anticancer drug. This study aimed to elucidate the impact of Fos APR on ISR in patients receiving paclitaxel and carboplatin, with and without bevacizumab therapy (TC±Bev). PATIENTS AND METHODS: This study focused on patients with gynecologic cancer (n=93) who received TC±Bev administration at Fujita Health University Hospital from March 2016 to February 2020, and monitored up to six cycles. The patients were randomly assigned to the Fos APR group (n=47) and the Aprepitant (APR) group (n=46). Using Visual Infusion Phlebitis (VIP) scores, ISR was evaluated by comparing the VIP scores of all cycles using a linear mixed model. The risk factors that contribute to the occurrence of vascular pain throughout all cycles were also examined. RESULTS: The VIP scores of all cycles showed a near significant intergroup difference (p=0.071). Factors that affected the development of vascular pain included Fos APR and age (p=0.027 and 0.049, respectively). Regarding age, patients aged <65 years had a higher risk. Four patients underwent a switch from the originally assigned neurokinin-1 receptor antagonist; in all of these cases, Fos APR was changed to APR for vascular pain. CONCLUSION: Fos APR may increase the risk for ISR associated with TC±Bev therapy for gynecological cancer.

Abstract Purpose In the context of in vitro fertilization–embryo transfer (IVF–ET), factors other than egg quality may be key determinants of treatment success, in particular, maternal factors related to uterine endometrial receptivity and unidentified factors. We therefore aimed to analyze the metabolome and microbiome in IVF–ET patients who did and did not achieve pregnancy. Methods Cervicovaginal mucus was collected from patients undergoing IVF–ET. Metabolite analysis was conducted by liquid chromatography‐mass spectrometry and the microbiota were determined by the polymerase chain reaction using universal 16S‐rRNA gene bacterial primers by MiSeq sequencing. Patients were classified as pregnant (N = 10) or nonpregnant (N = 13). Metabolic pathways were examined by MetaboAnalyst. Results Three metabolic pathways, including alanine‐aspartate–glutamate metabolism, arginine biosynthesis, and cysteine‐methionine metabolism, were commonly decreased at the time of embryo transfer irrespective pregnant outcomes. Notably, pyruvate was decreased in the pregnant group. Amino acid metabolites showed inverse correlations with the presence of anaerobic microbiota in the nonpregnant group. Conclusions Metabolism decreased during embryo transplantation, with a notable decrease in pyruvate metabolism, particularly in patients who became pregnant. The behavior of metabolites in the pregnant and nonpregnant groups suggests that metabolome analysis in the cervicovaginal mucus may be a diagnostic marker for predicting pregnancy.

Cervical cancer is the fourth most common cancer in women worldwide. Although cytology or HPV testing is available for screening, these techniques have their drawbacks and optimal screening methods are still being developed. Here, we sought to determine whether aberrant expression of miRNAs in cervical mucus could be an ancillary test for cervical neoplasms. The presence of miRNAs in 583 and 126 patients (validation and external cohorts) was determined by real-time RT-PCR. Performance of a combination with five miRNAs (miR-126-3p, -451a -144-3p, -20b-5p and -155-5p) was estimated by ROC curve analysis. Predicted probability (PP) was estimated by nomograms comprising -ΔCt values of the miRNAs, HPV genotype and age. A combination of five miRNAs showed a maximum AUC of 0.956 (95% CI: 0.933-0.980) for discriminating cancer. Low PP scores were associated with good prognosis over the 2-year observation period (p < 0.05). Accuracy for identifying cancer and cervical intraepithelial neoplasia (CIN) 3 + by nomogram was 0.983 and 0.966, respectively. PP was constant with different storage conditions of materials. We conclude that nomograms using miRNAs in mucus, HPV genotype and age could be useful as ancillary screening tests for cervical neoplasia.

エストロゲンにはさまざまな生理作用があり、食欲抑制はそのなかの一つである。近年、神経細胞内で生合成されるエストロゲン(ニューロエストロゲン)がさまざまな生理作用を持っていることが明らかになってきた。しかし、このニューロエストロゲンと食欲の関係は不明である。ニューロエストロゲンの食欲に対する作用を明らかにするため、卵巣切除したマウス(OVX)を作製して卵巣から分泌されるエストロゲンの影響を排除し、食欲とニューロエストロゲンの関係について検討した。その結果、OVXでは視床下部でのアロマターゼ発現量が増加し、食餌量は低下した。一方でOVXにアロマターゼ阻害薬(レトロゾール)を投与すると食餌量は増加した。このとき、視床下部での食欲抑制ペプチドPOMCの減少、食欲亢進ペプチドNpyの増加が観察された。さらに、アロマターゼを発現させたマウス由来視床下部神経細胞N38(TG)を用い、in vitroでエストロゲン、テストステロンを加えて検討した結果、N38(TG)によって合成されたエストロゲンによりPOMCの受容体であるMC4Rや、レプチン受容体が増加した。本研究では、卵巣から分泌されるエストロゲンではなく、ニューロエストロゲンによって食欲がコントロールされている可能性を示す。(著者抄録)

Persistent HPV infection associated with immune modulation may result in high-grade squamous intraepithelial lesions (CIN)2/3. Currently, there is little information on the cervicovaginal microbiome, local cytokine levels and HPV infection related to CIN. Follow-up of patients after local surgery provides an opportunity to monitor changes in the cervicovaginal environment. Accordingly, we undertook this longitudinal retrospective study to determine associations between HPV genotypes, cervicovaginal microbiome and local cytokine profiles in 41 Japanese patients with CIN. Cervicovaginal microbiota were identified using universal 16S rRNA gene (rDNA) bacterial primers for the V3/4 region by PCR of genomic DNA, followed by MiSeq sequencing. We found that Atopobium vaginae was significantly decreased (p < 0.047), whereas A. ureaplasma (p < 0.022) increased after surgery. Cytokine levels in cervical mucus were measured by multiplexed bead-based immunoassays, revealing that IL-1β (p < 0.006), TNF-α (p < 0.004), MIP-1α (p < 0.045) and eotaxin (p < 0.003) were significantly decreased after surgery. Notably, the level of eotaxin decreased in parallel with HPV clearance after surgery (p < 0.028). Thus, local surgery affected the cervicovaginal microbiome, status of HPV infection and immune response. Changes to the cervicovaginal microbiota and cervical cytokine profile following surgery for cervical intraepithelial neoplasia may be important for understanding the pathogenesis of CIN in future.

【目的】産婦人科良性疾患に対するロボット支援手術は、2018年4月より子宮全摘出術が新たに保険収載され、今後は全国的な普及が期待されているが、従来の腹腔鏡手術に対する臨床的な有効性は明らかとなっていない。そこで、産婦人科良性疾患に対するロボット支援下子宮全摘出術が保険収載された以降の2年間における従来の腹腔鏡手術とロボット支援手術の臨床成績について比較検討することを目的とした。【対象と方法】2018年4月より2020年3月に婦人科良性子宮疾患に対して従来の腹腔鏡下子宮全摘出術(conventional-laparoscopic hysterectomy:CLH)を施行した81例(CLH群)とロボット支援下子宮全摘出術(Robot assisted hysterectomy:RAH)を施行した62例(RAH群)を対象とした。検討項目は、両術式における子宮摘出重量、術中出血量、全手術時間、術後WBC、CRP(1日目、3日目)を後方視的に比較した。統計学的解析はマン・ホイットニーのU検定で行い、P<0.05を有意差ありと判定した。【結果】CLH群とRAH群を後方視的に比較検討した結果、年齢やBMI、子宮摘出重量、手術時間に差を認めなかったが、術中出血量(ml)は36(4-1,431) vs 30(5-306)、術後3日目のWBC(103/μL)は5.8(3.0-10.3) vs 5.1(3.0-10.1)とRAH群で低下する傾向があり(各p=0.07、p=0.09)、術後1日目のCRP(mg/dL)は3.5(0.4-10.9) vs 2.6(0.7-6.8)、術後3日目のCRP(mg/dL)は2.2(0.2-9.0) vs 1.8(0.2-5.5)とRAH群で有意な低下を認めた(各p=0.01、p=0.04)。【結語】当施設における婦人科良性子宮疾患に対するRAHは、CLHに比べ術中出血量や術後炎症反応が低下することが示されたことより、ロボット支援手術は手術侵襲を低減させる術式であると考えられた。(著者抄録)

Everolimus (EVE), an inhibitor of mammalian target of rapamycin, is an emerging second-line therapeutic option for hormone therapy-resistant breast cancers. However, some patients do not respond to EVE, whereas in others it exacerbates the disease. Cellular inhibitor of protein phosphatase 2A (CIP2A) is a human oncoprotein that can promote cancer cell growth and apoptosis resistance. Although CIP2A is upregulated in hormone-related cancers, such as breast cancer, little is known about potential anti-tumour effects of downregulating CIP2A. As a model to study the resistance of breast cancer cells to hormone treatment, we previously established clones of long-term estrogen depletion-resistant MCF-7 (LTED) cells. Here, we selected three clones highly responsive to EVE and three clones poorly responsive to EVE. When cells were treated with EVE, CIP2A mRNA expression was decreased in highly responsive EVE clones (DC-cells) whereas it was increased in poorly responsive EVE clones (IC-cells). Using Kaplan-Meier survival plots we report that high expression of CIP2A was associated with significantly reduced overall survival in patients with luminal A breast cancer. In IC-cells, cell growth was enhanced upon EVE treatment whereas an EVE range of 0.1-100 nM decreased growth in DC-cells. The mRNA expression of genes involved in epithelial-mesenchymal transition (EMT) such as CDH1, CLDN3, CK19 was significantly decreased in IC-cells, but remained unchanged in DC-cells. These findings highlight a relationship between CIP2A and EMT in the intrinsic resistance of hormone therapy-resistant breast cancers to EVE.