研究者詳細 - 小山内　誠

子宮頸部腺癌におけるclaudin-6発現とその意義

伊藤祐衣, 伊藤祐衣, 高澤啓, 車野晃大, 車野晃大, 桐澤くらら, 桐澤くらら, 高澤久美, 青山智志, 青山智志, 小野祐輔, 及能大輔, 小山内誠

日本病理学会会誌 110 ( 2 ) 2021年

血管内皮細胞のVEGFR2を介した肝蛍光発色技術の開発

及能大輔, 及能大輔, 高澤啓, 真柄和史, 仲盛優菜, 小野佑輔, 高澤久美, 竹政伊知朗, 小山内誠

日本病理学会会誌 110 ( 1 ) 2021年

乳癌におけるALDOAの発現とその病理学的意義

桐澤くらら, 桐澤くらら, 高澤啓, 車野晃大, 車野晃大, 伊藤祐衣, 伊藤祐衣, 高澤久美, 青山智志, 青山智志, 小野祐輔, 及能大輔, 小山内誠

日本病理学会会誌 110 ( 2 ) 2021年

レチノイン酸代謝酵素CYP26A1の発現は,乳がん組織のホルモンレセプター,HER2発現様式と関連する

小野佑輔, 山本夏子, 高澤久美, 真柄和史, 青山智志, 及能大輔, 高澤啓, 小山内誠

日本病理学会会誌 110 ( 1 ) 2021年

乳癌におけるJAM-Aの発現とその意義

真柄和史, 高澤啓, 高澤久美, 青山智志, 小野佑輔, 及能大輔, 小山内誠

日本病理学会会誌 110 ( 1 ) 2021年

胆道癌におけるタイト結合分子claudin18の発現状態を利用した病理学的診断および根治切除後の予後予測への有用性

及能大輔, 木村康利, 高澤啓, 計良淑子, 太田盛道, 永山稔, 今村将史, 西舘敏彦, 沖田憲司, 信岡隆幸, 小山内誠, 竹政伊知朗

日本外科学会定期学術集会(Web) 121st 2021年

乳癌におけるレチノイン酸代謝酵素CYP26A1発現の病理組織学的検討

山本夏子, 小野佑輔, 青山智志, 大門史士, 高澤久美, 高澤啓, 小山内誠

日本病理学会会誌 ( (一社)日本病理学会 ) 109 ( 1 ) 494 - 494 2020年03月

乳癌におけるALDOAの発現とその病理学的意義

桐澤くらら, 高澤啓, 伊藤祐衣, 車野晃大, 小野佑輔, 青山智志, 高澤久美, 小山内誠

日本病理学会会誌 ( (一社)日本病理学会 ) 109 ( 1 ) 494 - 494 2020年03月

子宮頸部腺がんにおける間質反応の臨床病理学的意義

高澤啓, 青山智志, 小野佑輔, 高澤久美, 村田雅樹, 小山内誠

日本病理学会会誌 ( (一社)日本病理学会 ) 109 ( 1 ) 370 - 370 2020年03月

子宮頸部腺癌におけるタイト結合関連タンパク質claudin-6発現とその意義

伊藤祐衣, 高澤啓, 桐澤くらら, 車野晃大, 青山智志, 小野佑輔, 高澤久美, 小山内誠

日本病理学会会誌 ( (一社)日本病理学会 ) 109 ( 1 ) 497 - 497 2020年03月

子宮頸部腺癌におけるタイト結合関連タンパク質JAM-Aの高発現は、癌悪性化に寄与する

村上太郎, 高澤啓, 青山智志, 小野佑輔, 高澤久美, 村田雅樹, 小山内誠

日本病理学会会誌 ( (一社)日本病理学会 ) 109 ( 1 ) 313 - 314 2020年03月

悪性末梢神経鞘腫のFFPE組織標本を用いた新規バイオマーカー探索の試み

車野晃大, 伊藤祐衣, 高澤啓, 桐澤くらら, 青山智志, 小野佑輔, 高澤久美, 小山内誠

日本病理学会会誌 ( (一社)日本病理学会 ) 109 ( 1 ) 498 - 498 2020年03月

子宮頸部腺がんで高発現するALDOAはがん悪性化に関与する

高澤啓, 高澤久美, 及能大輔, 小山内誠

日本癌学会学術総会抄録集(Web) 79th 2020年

子宮頸部腺癌細胞で高発現するclaudin-1のタイト結合機能に関する役割解析

高澤久美, 高澤啓, 青山智志, 北嶋洋志, 及能大輔, 小野佑輔, 村田雅樹, 小山内誠

日本分子生物学会年会プログラム・要旨集(Web) 43rd 2020年

FFPE組織を用いたプロテオミクスによる脂肪肉腫診断マーカーの探索

子宮頸部腺がんにおけるエストロゲン/GPR30を介したがん悪性化機構(Estrogen/GPR30 contributes to malignant potentials of uterine cervical adenocarcinoma via claudin-1 expression)

高澤啓, 高澤久美, 小山内誠

日本癌学会総会記事 ( (一社)日本癌学会 ) 78回 P - 1153 2019年09月

青山智志, 高澤啓, 小野祐輔, 長谷川匡, 小山内誠

JSBMS Letters ( (一社)日本医用マススペクトル学会 ) 44 ( Suppl. ) 106 - 106 2019年08月

Vasculogenic mimicryが見られたGIST(gastrointestinal stromal tumor)の一例

佐藤みどり, 小山内誠, 小野佑輔, 高澤啓, 村田雅樹, 三浦秀元, 秦史壯

日本病理学会会誌 ( (一社)日本病理学会 ) 108 ( 1 ) 451 - 451 2019年04月

タイト結合膜蛋白occludinはジスルフィド結合とHIF-1、ユビキチン化により安定性が制御される

田中敏, 小野佑輔, 高澤啓, 村田雅樹, 小山内誠, 澤田典均

日本病理学会会誌 ( (一社)日本病理学会 ) 108 ( 1 ) 331 - 331 2019年04月

タイト結合関連分子claudin-18は、レチノイン酸欠乏を原因とする乳がん悪性化に関与する

小野佑輔, 小山内誠, 青山智志, 高澤久美, 高澤啓, 村田雅樹

日本病理学会会誌 ( (一社)日本病理学会 ) 108 ( 1 ) 289 - 289 2019年04月

タイト結合関連タンパク質JAM-Aは子宮頸部腺癌で高発現し、癌悪性化に寄与する

村上太郎, 高澤啓, 青山智志, 小野佑輔, 高澤久美, 村田雅樹, 小山内誠

日本病理学会会誌 ( (一社)日本病理学会 ) 108 ( 1 ) 460 - 460 2019年04月

子宮頸部腺がんにおけるエストロゲン/GPR30/claudin-1を介したがん悪性化機構の解明(Estrogen/GPR30 contributes to malignant potentials of cervical adenocarcinoma via claudin-1)

高澤啓, 青山智志, 高澤久美, 小野佑輔, 村田雅樹, 澤田典均, 小山内誠

日本病理学会会誌 ( (一社)日本病理学会 ) 108 ( 1 ) 306 - 306 2019年04月

FFPE組織を用いた比較プロテオミクスによる高分化型脂肪肉腫のバイオマーカー探索(Identification of potential immunohistochemical markers for liposarcoma based on proteomic analysis using FFPE tissue)

高澤啓, 長谷川匡, 小山内誠

日本癌学会総会記事 ( (一社)日本癌学会 ) 77回 2257 - 2257 2018年09月

レチノイン酸代謝を起点として理解する細胞ががん化する仕組み

小山内誠, 小野佑輔, 高澤啓, 高澤久美, 村田雅樹, 澤田典均

日本病理学会会誌 ( (一社)日本病理学会 ) 107 ( 1 ) 340 - 340 2018年04月

子宮頸部腺癌におけるALDOA発現はその悪性形質に関与する

齋藤裕己, 高澤啓, 青山智志, 高澤久美, 小野佑輔, 村田雅樹, 小山内誠, 澤田典均

日本病理学会会誌 ( (一社)日本病理学会 ) 107 ( 1 ) 338 - 338 2018年04月

タイト結合分子JAM-Aの異常発現が肺腺癌の悪性化に関与する

真柄和史, 高澤啓, 青山智志, 太田未咲, 小野佑輔, 高澤久美, 村田雅樹, 小山内誠, 長谷川匡, 澤田典均

日本病理学会会誌 ( (一社)日本病理学会 ) 107 ( 1 ) 293 - 293 2018年04月

子宮頸部腺癌におけるJAM-A発現とその意義

村上太郎, 高澤啓, 齋藤裕己, 青山智志, 小野佑輔, 村田雅樹, 小山内誠, 澤田典均

日本病理学会会誌 ( (一社)日本病理学会 ) 107 ( 1 ) 526 - 526 2018年04月

claudin-18はEGFR/ERKシグナル経路と協働して胆道癌の悪性化に関与する(Overexpression of claudin-18 contributes to malignant potentials of bile duct cancer)

高澤久美, 高澤啓, 小山内誠, 青山智志, 小野佑輔, 村田雅樹, 澤田典均

日本病理学会会誌 ( (一社)日本病理学会 ) 107 ( 1 ) 342 - 342 2018年04月

FFPE組織を用いたプロテオミクスによる脂肪肉腫診断マーカーの探索

青山智志, 高澤啓, 村田雅樹, 小山内誠, 長谷川匡, 澤田典均

日本病理学会会誌 ( (一社)日本病理学会 ) 107 ( 1 ) 291 - 291 2018年04月

タイト結合膜蛋白occludinはジスルフィド結合とユビキチン化を介して安定性が調節され、アポトーシス誘導を制御する

田中敏, 高澤啓, 村田雅樹, 小山内誠, 澤田典均

生命科学系学会合同年次大会 ( 生命科学系学会合同年次大会運営事務局 ) 2017年度 [3P - 0144] 2017年12月

claudin-18はEGFR/ERK経路を介して胆道癌悪性化に関与する

高澤啓, 小山内誠, 澤田典均

日本癌学会総会記事 ( (一社)日本癌学会 ) 76回 J - 3050 2017年09月

平滑筋肉腫のFFPE組織標本を用いた新規バイオマーカー探索の試み

青山智志, 高澤啓, 伊野善彦, 小野佑輔, 村田雅樹, 小山内誠, 長谷川匡, 澤田典均

JSBMS Letters ( (一社)日本医用マススペクトル学会 ) 42 ( Suppl. ) 49 - 49 2017年08月

子宮頸部腺癌におけるALDOAの発現とその臨床病理学的意義

齋藤裕己, 高澤啓, 上田朝子, 太田未咲, 伊野善彦, 青山智志, 村田雅樹, 小山内誠, 澤田典均

日本病理学会会誌 ( (一社)日本病理学会 ) 106 ( 1 ) 511 - 511 2017年03月

慢性膿胸を背景に発症した両側副腎原発NK/T細胞性リンパ腫の一剖検例

河田由香, 高澤啓, 塚原智英, 村田雅樹, 小山内誠, 長谷川匡, 澤田典均

日本病理学会会誌 ( (一社)日本病理学会 ) 106 ( 1 ) 516 - 516 2017年03月

ランゲルハンス細胞肉腫を含む五重癌の一例

山本晃匡, 村田雅樹, 高澤啓, 小山内誠, 長谷川匡, 澤田典均

日本病理学会会誌 ( (一社)日本病理学会 ) 106 ( 1 ) 514 - 515 2017年03月

平滑筋肉腫のFFPE組織標本を用いた新規バイオマーカー探索の試み

高澤啓, 伊野善彦, 青山智志, 秋元太志, 中西敬太郎, 村田雅樹, 小山内誠, 長谷川匡, 澤田典均

日本病理学会会誌 ( (一社)日本病理学会 ) 106 ( 1 ) 288 - 288 2017年03月

子宮頸部腺癌における膜型Estrogen受容体GPR30の発現解析

伊野善彦, 高澤啓, 青山智志, 村田雅樹, 小山内誠, 長谷川匡, 澤田典均

日本病理学会会誌 ( (一社)日本病理学会 ) 106 ( 1 ) 343 - 343 2017年03月

乳癌におけるタイト結合関連タンパク質LSRの発現とその意義

太田未咲, 高澤啓, 上田朝子, 青山智志, 村田雅樹, 小山内誠, 長谷川匡, 澤田典均

日本病理学会会誌 ( (一社)日本病理学会 ) 106 ( 1 ) 510 - 510 2017年03月

原因蛋白質同定に難渋した心アミロイドーシスの1剖検例

上田朝子, 高澤啓, 太田未咲, 青山智志, 村田雅樹, 小山内誠, 澤田典均

日本病理学会会誌 ( (一社)日本病理学会 ) 106 ( 1 ) 507 - 507 2017年03月

Claudins in cancer: bench to bedside

Makoto Osanai, Akira Takasawa, Masaki Murata, Norimasa Sawada

PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY ( SPRINGER ) 469 ( 1 ) 55 - 67 2017年01月

書評論文，書評，文献紹介等

The claudin family, in mammals, encoded by at least 27 members of a single ancestral gene, CLDN, is the main constituent as integral membrane proteins of tight junctions. It has been shown that the expression levels of claudins are often decreased or that their expressions are absent in human neoplasias. These findings are consistent with the well-accepted concept that carcinogenesis is accompanied by the disruption or loss of functional tight junctions. In contrast, accumulating data have showed elevated or aberrant expression of claudins in various cancers, indicating specific roles of claudins in tumorigenesis. Importantly, dysregulated claudins play an oncogenic role or conversely have a tumor-suppressive effect depending on target tissues or cell types, and thus, they contribute to tumor development and progression. Although tight junctions are intercellular structures in epithelial cells, specific roles of claudins in cancer are supported by the evidence that TJs are not simple static constituents for establishing cell adhesion structures but are also cell signaling components that have functions in receiving environmental cues and transmitting signals inside cells. Since the expression profile of claudins is associated with patients' outcome and prognosis in several cancer types, an understanding of the expression pattern and subcellular localization of claudins in various pathologies will lead to the establishment of claudins as useful biomarkers for the detection and diagnosis of cancers.

肺腺癌におけるJAM-Aの発現とその意義

真柄和史, 高澤啓, 村田雅樹, 田上洋平, 秋元太志, 田中敏, 小山内誠, 長谷川匡, 澤田典均

日本病理学会会誌 ( (一社)日本病理学会 ) 105 ( 1 ) 588 - 588 2016年04月

claudin-1は子宮頸部腺癌細胞の浸潤に関与し、claudin-1の発現はエストロゲンにより調節される

秋元太志, 高澤啓, 村田雅樹, 上田朝子, 青山智志, 田中敏, 小山内誠, 澤田典均

日本病理学会会誌 ( (一社)日本病理学会 ) 105 ( 1 ) 349 - 349 2016年04月

平滑筋肉腫のFFPE組織標本を用いた新規バイオマーカー探索の試み

中西敬太郎, 高澤啓, 青山智志, 上田朝子, 秋元太志, 村田雅樹, 田中敏, 小山内誠, 長谷川匡, 澤田典均

日本病理学会会誌 ( (一社)日本病理学会 ) 105 ( 1 ) 602 - 602 2016年04月

FFPE組織を用いた比較プロテオミクスによる高分化型脂肪肉腫のバイオマーカー探索

青山智志, 高澤啓, 村田雅樹, 中西敬太郎, 田中敏, 小山内誠, 長谷川匡, 澤田典均

日本病理学会会誌 ( (一社)日本病理学会 ) 105 ( 1 ) 338 - 338 2016年04月

レチノイン酸代謝酵素CYP26A1は、fascinの発現増加を介して乳がんの進展に関与する

小山内誠, 李康弘, 高澤啓, 村田雅樹, 田中敏, 澤田典均

日本病理学会会誌 ( (一社)日本病理学会 ) 105 ( 1 ) 471 - 471 2016年04月

子宮頸部腺癌におけるタイト結合関連たんぱく質の免疫組織化学的検討

高澤啓, 秋元太志, 村田雅樹, 青山智志, 田中敏, 小山内誠, 長谷川匡, 澤田典均

日本病理学会会誌 ( (一社)日本病理学会 ) 105 ( 1 ) 478 - 478 2016年04月

子宮頸部腺癌における受容体型チロシンキナーゼの発現解析

上田朝子, 高澤啓, 秋元太志, 中西敬太郎, 村田雅樹, 青山智志, 田中敏, 小山内誠, 長谷川匡, 澤田典均

日本病理学会会誌 ( (一社)日本病理学会 ) 105 ( 1 ) 602 - 602 2016年04月

Elevated expression of the retinoic acid-metabolizing enzyme CYP26C1 in primary breast carcinomas

Makoto Osanai, Gang-Hong Lee

MEDICAL MOLECULAR MORPHOLOGY ( SPRINGER JAPAN KK ) 49 ( 1 ) 22 - 27 2016年03月

Retinoic acid (RA)-metabolizing enzyme CYP26A1 has been shown to have increased expression levels in breast cancers and to effectively promote the survival of breast carcinoma cells, implying a potential oncogenic function. However, the expression of CYP26C1, another CYP26 family member, in primary breast carcinoma remains to be clarified. In the present study, we examined the expression of CYP26C1 by immunohistochemistry, using three different types of microarray, and observed strong cytoplasmic staining of CYP26C1 in 73 of the 219 (33.3 %) breast carcinomas. In contrast, CYP26C1 was not expressed in normal ductal and lobular cells in non-neoplastic tissue. Interestingly, increased expression of CYP26C1 was significantly associated with a high Ki-67 labeling index and a grade of tumor. However, CYP26C1 immunoreactivity was not associated with clinicopathological variables, including primary tumor status, lymph node involvement, distant metastasis, and tumor stage. In addition, CYP26C1 positivity was independent of the expression status of the hormone receptors and immunohistochemical surrogates for the intrinsic subtypes of breast cancer. This report is the first to demonstrate elevated expression of CYP26C1 in primary breast carcinomas. Based on the RA-catabolizing activity of CYP26C1, our data suggest that CYP26C1 expression may contribute to neoplasia in the breast.

Vasculogenic mimicry in gastrointestinal stromal tumor of the stomach; A case report

Osanai M, Takasawa A, Murata M, Tanaka S, Miura H, Hata F, Sawada N

Case Rep Clin Pathol 3 50 - 54 2016年 [査読有り]

タイト結合蛋白occludinの低酸素下でのジスルフィド結合とユビキチン化を介した細胞内分布調節

田中敏, 小野佑輔, 高澤啓, 村田雅樹, 高澤久美, 小山内誠, 澤田典均

日本生化学会大会・日本分子生物学会年会合同大会講演要旨集 ( (公社)日本生化学会 ) 88回・38回 [2P0424] - [2P0424] 2015年12月

The retinoic acid-metabolizing enzyme CYP26A1 upregulates fascin and promotes the malignant behavior of breast carcinoma cells

Makoto Osanai, Gang-Hong Lee

ONCOLOGY REPORTS ( SPANDIDOS PUBL LTD ) 34 ( 2 ) 850 - 858 2015年08月 [査読有り]

The retinoic acid (RA)-metabolizing enzyme CYP26A1 has been shown to efficiently enhance the oncogenic potential of breast cancer, suggesting a potential oncogenic function. We previously demonstrated that CYP26A1 confers unique cell survival properties by modulating the expression of a variety of genes and identified a number of genes that drive the cells into the oncogenic state. Accumulating evidence suggested that fascin is overexpressed in various types of cancer, primarily leading to increased cell motility. Therefore, in the present study, we examined fascin, an actin-bundling protein, using immunohistochemical and SA-P-gal staining as well as TUNEL and colony forming assays. The results of the present study showed that the expression levels of fascin increased significantly in response to CYP26A1 overexpression and, conversely, treatment with all-trans RA downregulated the expression of fascin. In addition, primary breast carcinoma samples, particularly hormone receptor-negative carcinomas and CYP26A1-overexpressing cancers, expressed elevated levels of fascin. Notably, fascin contributed to the ability of breast carcinoma cells to escape premature senescence and exhibit enhanced cell apoptotic resistance, promoting anchorage-independent growth properties. Fascin also promoted cell motility and the invasiveness of CYP26A1-expressing breast carcinoma cells. These data suggest that fascin expression is modulated by the intracellular RA status regulated by the expression of CYP26A1 and plays a significant role in the malignant behavior of CYP26A1-expressing breast carcinoma cells. CYP26A1 exerts oncogenic functions during breast carcinogenesis. Therefore, CYP26A1-mediated oncogenic characteristics may be partially responsible for the elevated expression of fascin.

Increased expression of the retinoic acid-metabolizing enzyme CYP26A1 during the progression of cervical squamous neoplasia and head and neck cancer

Makoto Osanai, Gang-Hong Lee

BMC Research Notes ( BioMed Central Ltd. ) 7 ( 1 ) 697 - 697 2014年10月 [査読有り]

Background: Retinoic acid (RA) is a critical regulator of cell differentiation, proliferation, and apoptosis in various cell types. Recently, the RA-metabolizing enzyme CYP26A1 (cytochrome P450, family 26, subfamily A, polypeptide 1) has been shown to have an oncogenic function in breast carcinogenesis. However, the relevance of elevated CYP26A1 expression in human cancers remains to be clarified. Methods. We immunohistochemically examined the expression of CYP26A1 in cervical squamous cell carcinoma (SCC) and its precursors, including low- and high-grade squamous intraepithelial lesions (LSIL and HSIL, respectively), as well as head and neck cancer (HNC). The association between CYP26A1 expression and a number of clinicopathological parameters was also evaluated. Results: CYP26A1 was not expressed in normal cervical epithelium. CYP26A1 expression was present in LSIL but limited to basal and parabasal cells. HSIL cases exhibited strong nuclear expression of CYP26A1 and mixed cytoplasmic expression patterns with widely distributed expression toward the epithelial surface. Importantly, strong cytoplasmic staining of CYP26A1 was observed in 19 of 50 (38%) patients with cervical SCC. Elevated expression of CYP26A1 was significantly associated with younger age (< 50 years) and lymph node involvement (pN). Similarly, CYP26A1 was not expressed in non-neoplastic tissues of the head and neck, but strong cytoplasmic staining of CYP26A1 was observed in 52 of 128 (41%) HNC cases. Such strong CYP26A1 expression was significantly associated with the primary tumor stage of carcinomas (pT) and the pathological tumor-node-metastasis (pTNM) stage in HNC. Conclusion: Our results indicated an elevated CYP26A1 expression in malignant and precancerous dysplastic lesions of the human cervix, which also increased with the progression of cervical squamous neoplasia. In addition, this report is the first to demonstrate the increased expression of CYP26A1 in HNC and its significant correlation with primary tumor growth. These data suggested that CYP26A1 overexpression might contribute to the development and progression of cervical malignancies and squamous neoplasia of the head and neck.

Enhanced expression of retinoic acid-metabolizing enzyme CYP26A1 in sunlight-damaged human skin

Makoto Osanai, Gang-Hong Lee

MEDICAL MOLECULAR MORPHOLOGY ( SPRINGER TOKYO ) 44 ( 4 ) 200 - 206 2011年12月

Vitamin A deficiency (VAD) is associated with increased susceptibility to carcinogenesis. CYP26A1, the gene encoding a cytochrome P450 enzyme specifically involved in metabolic inactivation of retinoic acid (RA), the most active vitamin A derivative, has been shown to result in a state of functional VAD of the cell. Recently, we demonstrated that CYP26A1 efficiently promotes cell survival properties and eventually contributes to the carcinogenic process, implying roles as an oncogene. To clarify the possible association between VAD caused by CYP26A1 expression and the development of human epithelial neoplasia, we examined whether enhanced expression of CYP26A1 might be observed in various lesions of human skin. We report here that basal keratinocytes showed only weak positivity of CYP26A1 in sunlight-nonexposed areas, whereas strong positive staining was observed in skin from chronically sunexposed body areas and in epidermis that had the dysplastic changes known as actinic keratosis. However, we found no expression of constitutive CYP26A1 in skin malignancies such as squamous cell carcinomas. Our observation suggests an involvement of enhanced CYP26A1 expression causing a functional VAD state in skin that can potentially lead to neoplastic transformation of keratinocytes in an early phase during skin carcinogenesis.

Oncogenic and cell survival properties of the retinoic acid metabolizing enzyme, CYP26A1

M. Osanai, N. Sawada, G-H Lee

ONCOGENE ( NATURE PUBLISHING GROUP ) 29 ( 8 ) 1135 - 1144 2010年02月

Vitamin A deficiency (VAD) is associated with increased susceptibility to carcinogenesis in animal models and elevated risk for a number of human cancers. Here, we found that CYP26A1, the gene encoding a cytochrome P450 enzyme specifically involved in metabolic inactivation of retinoic acid (RA), the most active vitamin A derivative, is highly expressed in 42% (27/65) of primary breast cancers. We also showed that enhanced expression of CYP26A1 suppresses cellular responses to anoikis and consequently promotes anchorage-independent growth. This transformed phenotype was sufficient to markedly increase tumorigenic and metastatic potential. Suppression of CYP26A1 significantly reversed the CYP26A1-mediated oncogenic characteristics, suggesting a direct link between intracellular RA status and tumorigenicity. Our observations provide strong evidence for oncogenic and cell survival properties of CYP26A1 in carcinogenesis, and suggest mechanisms whereby VAD might promote cancer development. Oncogene (2010) 29, 1135-1144; doi: 10.1038/onc.2009.414; published online 23 November 2009

Emerging novel treatment strategies for diabetic eye diseases

Makoto Osanai, Nami Nishikiori, Gang-Hong Lee, Norimasa Sawada

Current Diabetes Reviews 6 ( 1 ) 35 - 41 2010年01月

書評論文，書評，文献紹介等

Endothelial tight junctions (TJs) in the retina are potential therapeutic targets for diabetic complications such as retinopathy. TJs primarily determine the endothelial barrier, regulating vascular permeability to maintain tightly closed circulating homeostasis. Our recent study has demonstrated that glial cell-derived cytokines limit vascular permeability by modulating the TJ function of retinal capillary endothelium and eventually attenuate the breakdown of vascular integrity in diabetic microangiopathy. In addition, suppression of deregulated protease activity in lens cells results in dramatic inhibition of the development of murine diabetic cataracts. We believe that pharmacological modulation of the ocular tissue microenvironment such as that regulated by endothelial TJs may be a feasible strategy for reducing the development of diabetic complications in the eye. © 2010 Bentham Science Publishers Ltd.

Transmembrane proteins of tight junctions

Hideki Chiba, Makoto Osanai, Masaki Murata, Takashi Kojima, Norimasa Sawada

BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES ( ELSEVIER SCIENCE BV ) 1778 ( 3 ) 588 - 600 2008年03月

書評論文，書評，文献紹介等

Tight junctions contribute to the paracellular barrier, the fence dividing plasma membranes, and signal transduction, acting as a multifunctional complex in vertebrate epithelial and endothelial cells. The identification and characterization of the transmembrane proteins of tight junctions, claudins, junctional adhesion molecules (JAMs), occludin and tricellulin, have led to insights into the molecular nature of tight junctions. We provide an overview of recent progress in studies on these proteins and highlight their roles and regulation, as well as their functional significance in human diseases. (C) 2007 Elsevier B.V. All rights reserved.

Epigenetic silencing of claudin-6 promotes anchorage-independent growth of breast carcinoma cells

Makoto Osanai, Masaki Murata, Hideki Chiba, Takashi Kojima, Norimasa Sawada

CANCER SCIENCE ( BLACKWELL PUBLISHING ) 98 ( 10 ) 1557 - 1562 2007年10月

Cancer cells often exhibit loss of functional tight junctions (TJ), and disruption of the TJ structure is associated with cancer development. However, whether loss of a certain type of claudin, an integral membrane protein of TJ, is involved in malignant phenotypes remains to be clarified. Based on a report that claudin-6 functions as a tumor suppressor for breast cancer, the authors show here that suppression of claudin-6 expression results in increased resistance to various apoptogens, and causally enhances anchorage-independent growth properties. Because claudin-6 expression is partially silenced by promoter CpG island hypermethylation in MCF7 breast carcinoma cells, a synergistic effect of a demethylator and histone deacetylase inhibitor up-regulates the expression of endogenous claudin-6, which is sufficient for apoptotic sensitization and abrogation of colony-forming efficacy. In addition, decreased expression of claudin-6 promotes cellular invasiveness and transendothelial migration, accompanied by an increase in matrix metalloproteinase activity. These data suggest that the methylator phenotype of claudin-6 may at least partially contribute to enhanced tumorigenic and invasive properties of breast carcinoma cells.

Occludin-mediated premature senescence is a fail-safe mechanism against tumorigenesis in breast carcinoma cells

Makoto Osanai, Masaki Murata, Nami Nishikiori, Hideki Chiba, Takashi Kojima, Norimasa Sawada

CANCER SCIENCE ( BLACKWELL PUBLISHING ) 98 ( 7 ) 1027 - 1034 2007年07月

We have previously demonstrated that epigenetic silencing of occludin, a tight junction-associated membrane protein, results in the acquisition of apoptotic resistance to various apoptogenic stimuli, causally contributing to the enhanced tumorigenicity of cancer cells. However, it remains to be examined whether occludin expression in transformed cells has an alternative impact that is important for cancer progression. Here we show that forced expression of occludin induces anoikis and promotes oxidative stress-induced premature senescence in breast carcinoma cells, which is accompanied by upregulation of negative cell cycle regulators such as p16(INK4A), p21(Waf1/Cip1) and p27(Kip1) but not p53. The senescent phenotype is reversed by specific inhibition of mitogen-activated protein kinase. Endogenous reexpression of occludin mediated by a synergistic effect with a demethylator and histone deacetylase inhibitor or retinoids that stimulate retinoic acid receptor alpha is also sufficient for provoking the senescent phenotype. In addition, tumors that developed from occludin-expressing cells in mice showed a feature of cellular senescence that has not been described as a consequence of occludin signaling. These findings suggest that the loss of occludin expression is at least partially involved in the senescence-escape program during mammary tumorigenesis.

HNF4αによる上皮極性形成におけるタイト結合膜裏打ち分子cingulinの機能

杉本幸太郎, 千葉英樹, 酒井直行, 小山内誠, 小島隆, 澤田典均

日本病理学会会誌 ( (一社)日本病理学会 ) 96 ( 1 ) 358 - 358 2007年02月

Cellular retinoic acid bioavailability determines epithelial integrity: Role of retinoic acid receptor alpha agonists in colitis

Makoto Osanai, Nami Nishikiori, Masaki Murata, Hideki Chiba, Takashi Kojima, Norimasa Sawada

MOLECULAR PHARMACOLOGY ( AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS ) 71 ( 1 ) 250 - 258 2007年01月

The epithelial barrier is determined primarily by intercellular tight junctions (TJs). We have demonstrated previously that all-trans retinoic acid (atRA) plays an important role in forming functional TJs through a specific retinoic acid receptor (RAR)/retinoid X receptor heterodimer in epithelial cells. However, the physiological relevance of retinoic acids (RAs) in maintaining the epithelial integrity remains to be examined. Here, we show that several types of RA, including atRA, promote the barrier function of epithelial TJs. Conversely, RA depletion in the cells by overexpressing CYP26s, cytochrome P450 enzymes specifically involved in the metabolic inactivation of RAs, induces an increase of permeability as measured by two differently sized tracer molecules, inulin and mannitol. This RA-mediated enhancement of barrier function is potentially associated with the increased expression of TJ-associated genes such as occludin, claudin-1, claudin-4, and zonula occludens-1. We also found that RAR alpha is a preferential regulator of the epithelial barrier in vitro. Studies of murine experimental colitis, which is characterized by increased gut permeability, reveal that RAR alpha stimulation significantly attenuates the loss of the epithelial barrier during colitis in vivo. Our results suggest that cellular RA bioavailability determines the epithelial integrity, because it is a critical regulator for barrier protection during mucosal injuries.

Epigenetic silencing of occludin promotes tumorigenic and metastatic properties of cancer cells via modulations of unique sets of apoptosis-associated genes

Makoto Osanai, Masaki Murata, Nami Nishikiori, Hideki Chiba, Takashi Kojima, Norimasa Sawada

CANCER RESEARCH ( AMER ASSOC CANCER RESEARCH ) 66 ( 18 ) 9125 - 9133 2006年09月

Occludin is the first identified integral protein for the tight junction (TJ), and its long COOH-terminal domain is considered to have functions in receiving and transmitting cell survival signals. Loss of TJ-associated molecules, such as occludin, has been correlated with tumor progression in carcinogenesis; however, the precise molecular mechanisms explaining its loss of expression and whether occludin expression has any effects on cancer phenotypes remain to be clarified. Here, we show that forced expression of occludin in cancer cells exhibits enhanced sensitivity to differently acting apoptogenic factors, and thus inhibits the tumorigenicity of transformed cells, via modulation of unique sets of apoptosis-associated genes. In addition, studies using deletion mutants of occludin constructs show that 44 amino acids at the COOH-terminal end play a critical role in modifying the cellular phenotypes. Interestingly, occludin decreases cellular invasiveness and motility, thereby abrogating metastatic potencies of cancer cells. We also found that occludin expression is silenced by CpG island hypermethylation on its promoter region. Synergy with a demethylator and histone deacetylase inhibitor or retinoids that stimulate retinoic acid receptor alpha induces endogenous occludin, which is sufficient for apoptotic sensitization. Our results show the functional diversity of occludin and suggest that methylator phenotype of occludin provides enhanced tumorigenic, invasive, and metastatic properties of cancer cells, identifying occludin as a likely candidate for a tumor-suppressor gene in certain types of cancer.

HNF-4αはタイト結合膜裏打ち分子cingulinの発現を誘導する

杉本幸太郎, 千葉英樹, 藤田裕樹, 小山内誠, 小島隆, 澤田典均

日本病理学会会誌 ( (一社)日本病理学会 ) 95 ( 1 ) 390 - 390 2006年04月

Occludin expression inhibits tumorigenicity and metastasis

M Osanai, N Nishikiori, M Murata, H Chiba, T Kojima, N Sawada

FASEB JOURNAL ( FEDERATION AMER SOC EXP BIOL ) 20 ( 4 ) A223 - A223 2006年03月

研究発表ペーパー・要旨（国際会議）

Glial cell line-derived neurotrophic factor (GDNF) regulates the vascular permeability of the blood-retinal barrier (BRB)

N Nishikiori, M Osanai, H Miyajima, H Chiba, T Kojima, N Sawada

FASEB JOURNAL ( FEDERATION AMER SOC EXP BIOL ) 20 ( 4 ) A636 - A637 2006年03月

研究発表ペーパー・要旨（国際会議）

The significance of interferon-gamma-triggered internalization of tight-junction proteins in inflammatory bowel disease.

Hideki Chiba, Takashi Kojima, Makoto Osanai, Norimasa Sawada

Science's STKE : signal transduction knowledge environment 2006 ( 316 ) pe1 2006年

書評論文，書評，文献紹介等

Disruption of the epithelial barrier function of tight junctions by the proinflammatory cytokine interferon (IFN)-gamma plays a fundamental role in the pathogenesis of inflammatory bowel disease and other gastrointestinal disorders, but its precise mechanism has not been established. This Perspective provides an overview of how IFN-gamma triggers barrier dysfunction and bacterial translocation in intestinal epithelial cells, and highlights the roles of internalization of tight-junction transmembrane proteins, as well as rearrangements of the cortical actin cytoskeleton, in intestinal inflammation.

Behavior of tight-junction, adherens-junction and cell polarity proteins during HNF-4 alpha-induced epithelial polarization

S Satohisa, H Chiba, M Osanai, S Ohno, T Kojima, T Saito, N Sawada

EXPERIMENTAL CELL RESEARCH ( ELSEVIER INC ) 310 ( 1 ) 66 - 78 2005年10月

We previously reported that expression of tight-junction molecules occludin, claudin-6 and claudin-7, as well as establishment of epithelial polarity, was triggered in mouse F9 cells expressing hepatocyte nuclear factor (HNF)-4 alpha [H. Chiba, T. Gotoh, T. Kojima, S. Satohisa, K. Kikuchi, M. Osanai, N. Sawada. Hepatocyte nuclear factor (HNF)-4a triggers formation of functional tight junctions and establishment of polarized epithelial morphology in F9 embryonal carcinoma cells, Exp. Cell Res. 286 (2003) 288-297]. Using these cells, we examined in the present study behavior of tight-junction, adherens-junction and cell polarity proteins and elucidated the molecular mechanism behind HNF-4 alpha-initiated junction formation and epithelial polarization. We herein show that not only ZO-1 and ZO-2, but also ZO-3, junctional adhesion molecule (JAM)-B, JAM-C and cell polarity proteins PAR-3, PAR-6 and atypical protein kinase C (aPKC) accumulate at primordial adherens junctions in undifferentiated F9 cells. In contrast, CRB3, Pals1 and PATJ appeared to exhibit distinct subcellular localization in immature cells, Induced expression of HNF-4 alpha led to translocation of these tight-junction and cell polarity proteins to beltlike tight junctions, where occludin, claudin-6 and claudin-7 were assembled, in differentiated cells. Interestingly, PAR-6, aPKC, CRB3 and Pals1, but not PAR-3 or PATJ, were also concentrated on the apical membranes in differentiated cells. These findings indicate that HNF-4 alpha provokes not only expression of tight-junction adhesion molecules, but also modulation of subcellular distribution of junction and cell polarity proteins, resulting in junction formation and epithelial polarization. (c) 2005 Elsevier Inc. All rights reserved.

Down-regulation of survival signaling through MAPK and Akt in occludin-deficient mouse hepatocytes in vitro

M Murata, T Kojima, T Yamamoto, M Go, K Takano, M Osanai, H Chiba, N Sawada

EXPERIMENTAL CELL RESEARCH ( ELSEVIER INC ) 310 ( 1 ) 140 - 151 2005年10月

The tight junction (TJ) regulates epithelial cell polarity and barrier including permeability of the paracellular pathway. Occludin was the first integral membrane protein to be discovered, but it is not indispensable for the formation of TJ strands. The physiological function of occludin is still unclear, although occludin-deficient mice show very complex abnormalities in various organs without overt dysfunction of the TJ. To investigate the role of occludin in TJ expression and apoptosis regulated by survival signal transduction pathways such as MAPK and Akt, we performed primary culture of hepatocytes and established hepatic cell lines from occludin-deficient mice. In primary cultures of occludin-deficient mouse hepatocytes, claudin-2 expression and apoptosis were induced by down-regulation of the activation of MAPK and Akt. In them hepatic cell lines derived from occludin-deficient mice, claudin-2 expression and serum-free induced apoptosis were also increased by down-regulation of the activation of MAPK and Akt. Furthermore, in the hepatic cell lines transiently transfected with mouse and rat occludin genes, induction of claudin-2 expression and the apoptosis were inhibited with increases in activation of MAPK and Akt. These findings show that occludin plays a crucial role in claudin-2-dependent TJ function and the apoptosis involving MAPK and Akt signaling pathways in hepatocytes. (c) 2005 Elsevier Inc. All rights reserved.

HNF-4a provokes phosphorylation of CRB3 and develops junctional complexes and epithelial cell polarity

Hideki Chiba, Seiro Satohisa, Makoto Osanai, Shigeo Ohno, Takashi Kojima, Norimasa Sawada

CELL STRUCTURE AND FUNCTION ( JAPAN SOC CELL BIOLOGY ) 30 87 - 87 2005年06月

研究発表ペーパー・要旨（国際会議）

Glyceraldehyde-derived advanced glycation end-products preferentially induce VEGF expression and reduce GDNF expression in human astrocytes

H Miyajima, M Osanai, H Chiba, N Nishikiori, T Kojima, K Ohtsuka, N Sawada

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS ( ACADEMIC PRESS INC ELSEVIER SCIENCE ) 330 ( 2 ) 361 - 366 2005年05月

The blood-brain barrier (BBB) is a biological unit composed of capillary endothelial cells and astrocytes. Here we examined the effects of various types of advanced glycation end-products (AGEs) on astrocytes and BBB-forming endothelial cells. While no type of AGE we examined changed the permeability of endothelial sheets, glyceraldehyde-derived AGE induced VEGF expression most significantly in astrocytes. The expression of glial cell line-derived neurotrophic factor (GDNF), which reduces the vascular permeability, was decreased in the astrocytes by treatment with glyceraldehyde-derived AGE. These results indicate that glyceraldehyde-derived AGE is the biologically active substance for astrocytes by regulating the VEGF and GDNF expression, which is causally contributing to an increase in the permeability of the BBB. (c) 2005 Elsevier Inc. All rights reserved.

Expression of the retinoic acid-metabolizing enzyme CYP26A1 limits programmed cell death

M Osanai, M Petkovich

MOLECULAR PHARMACOLOGY ( AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS ) 67 ( 5 ) 1808 - 1817 2005年05月

Vitamin A deficiency has been associated with increased incidence of certain types of cancer; however, the mechanisms by which vitamin A depletion promotes tumorigenesis are poorly understood. In addition all-trans-retinoic acid (RA), the most active form of vitamin A metabolites, has been shown to limit carcinogenesis in animal models and to trigger programmed cell death (apoptosis) in certain types of tumor cells. On the other hand, we show here that various cell lines overexpressing CYP26A1, a cytochrome P450 enzyme specifically involved in the catabolic inactivation of RA, exhibit increased resistance to various apoptogenic factors, including death receptor ligands such as tumor necrosis factor-related apoptosis-inducing ligand. This resistance could be reversed by pretreatment with ketoconazole, a broad-spectrum inhibitor of cytochrome P450 enzymes. In addition, synthetic retinoids Am80 (4[( 5,6,7,8-tetrahydro5,5,8,8- tetramethyl-2-naphthalenyl) carbamoyl] benzoic acid) and Am580 [ 4( 5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphtamido) benzoic acid], which are resistant to CYP26A1 metabolism, can restore the sensitivity of these cells to apoptogens. Thus, these findings support the idea that CYP26 expression levels may play a role in determining cellular commitment to apoptosis, and increased RA metabolism may be at least partially responsible for these observed effects.

Phosphorylation of ezrin enhances microvillus length via a p38 MAP-kinase pathway in an immortalized mouse hepatic cell line

MD Lan, T Kojima, M Murata, M Osanai, K Takano, H Chiba, N Sawada

EXPERIMENTAL CELL RESEARCH ( ELSEVIER INC ) 312 ( 2 ) 111 - 120 2005年01月

The apical microvilli are closely related with the development and the maintenance of cell polarization, and the length of microvilli varies in a regular way among cell types. Ezrin, a member of the ezrin/radixin/moesin (ERM) family, seems to be involved in the formation and stabilization of the apical microvilli. We found that phosphorylation of ezrin caused elongation of microvilli via a p38 MAP-kinase signaling pathway in all immortalized mouse hepatic cell line. When, in the oncogenic Raf-1-transfected mouse hepatic cell line, epithelial to mesenchymal transition (EMT) indicated as down-regulation of E-cadherin and up-regulation of Snail occurred, loss of microvilli and down-regulation of ezrin hot not radixin and moesin were also observed. In the Raf-1 transfectants treated with the MAP-kinase inhibitor PD98059 and the p38 MAP-kinase inhibitor SB203580, the numbers of microvilli and the expression of ezrin, E-cadherin and Snail were recovered. More interestingly, treatment with SB203580 induced elongation of microvilli and increased phosphorylation of ezrin (at Thr-567 and Tyr-353). Phosphorylated ezrin-positive clots were colocalized with actin-positive dots on the surface of some Raf-1 transfectants treated with SB203580. These results suggested that phosphorylation of ezrin via the p38 MAP-kinase signaling pathway might be involved in the formation of microvilli during development of epithelial cell polarization. (c) 2005 Elsevier Inc. All rights reserved.

Activation of p21 (CIP1/WAF1) gene expression and inhibition of cell proliferation by overexpression of hepatocyte nuclear factor-4 alpha

H Chiba, T Itoh, S Satohisa, N Sakai, H Noguchi, M Osanai, T Kojima, N Sawada

EXPERIMENTAL CELL RESEARCH ( ELSEVIER INC ) 302 ( 1 ) 11 - 21 2005年01月

The F9 murine embryonal carcinoma cell line provides an attractive system for studying epithelial differentiation and antiproliferative processes. We have recently established F9 cells expressing doxycycline-inducible hepatocyte nuclear factor (HNF)-4alpha and shown that HNF-4alpha triggers the gene expression of tight-junction molecules, occludin, claudin-6, and claudin-7, as well as formation of functional tight junctions and polarized epithelial morphology (Exp. Cell Res. 286, [2003] 288). Since these events were very similar to those induced by retinoids, we investigated whether HNF-4alpha, like retinoid receptors, was involved in the control of cell proliferation. We herein show that HNF-4alpha up-regulates expression of the p21 gene, but not the p15, p16, p18, p19, or p27 gene, in a p53-independent manner, and inhibits cell growth in F9 cells. Similar results were observed in rat lung endothelial cells, in which expression of HNF-4alpha is conditionally induced by doxycycline. Furthermore, we demonstrate, by reporter assay, that HNF-4alpha significantly elevates the transcriptional activity of the p21 promoter. Since, HNF-4alpha is expressed not only in the liver but also in organs containing epithelial cells, such as kidney, intestine, pancreas, and stomach, it might also play critical roles in the regulation of epithelial morphogenesis and proliferation in these organs. (C) 2004 Elsevier Inc, All rights reserved.

Oncogenic Raf-1 regulates epithelial to mesenchymal transition via distinct signal transduction pathways in an immortalized mouse hepatic cell line

MD Lan, T Kojima, M Osanai, H Chiba, N Sawada

CARCINOGENESIS ( OXFORD UNIV PRESS ) 25 ( 12 ) 2385 - 2395 2004年12月

The epithelial to mesenchymal transition (EMT) is considered to be an important event during malignant tumor progression and metastasis. Although Raf/MEK/ERK signaling causes EMT, the mechanisms, including the signaling pathways, are as yet unclear. In the present study we have examined the effects of signal transduction pathways on oncogenic Raf-1-induced EMT, using an immortalized mouse hepatic cell line. Oncogenic Raf-1-induced EMT is characterized by down-regulation of adherens and tight junctions and the reorganization of actin. An active Raf-1 gene was introduced into a mouse hepatic cell line which was then treated with the MAP kinase inhibitor PD98059, the p38 MAP kinase inhibitor SB203580, the PI3 kinase inhibitor LY294002 or the c-Src tyrosine kinase inhibitor PP2. The expression and localization of the adherens and tight junction proteins E-cadherin, occludin, ZO-1, claudin-1 and claudin-2 were determined by western blotting, RT-PCR and immunocytochemistry. The barrier function of tight junctions was assessed by measurements of transepithelial electric resistance (TER) and permeability in terms of fluxes of [C-14]mannitol and [C-14]inulin. In Raf-1-transfected cells expression of occludin and claudin-2 was markedly down-regulated at the protein and mRNA levels and the TER value was decreased, while the permeability was increased. The distribution of ZO-1, pancadherin and F-actin was changed from linear to zipper-like structures at cell borders. In Raf-1-transfected cells treated with PD98059 and SB203580, but not LY294002, expression and localization of claudin-2, but not occludin, recovered, together with barrier function, measured as the TER value. The distributions of ZO-1, pancadherin and F-actin also recovered on treatment with PD98059 and SB203580, but not LY294002. Expression and localization of occludin recovered slightly on treatment with PP2. Thus, oncogenic Raf-1 regulates EMT via distinct MAP kinase, p38 MAP kinase and c-Src tyrosine kinase signal pathways in the mouse hepatic cell line.

Unique cellular features of peripheral primitive neuroectodermal tumor: Ultrastructural evidence of its unique cytodifferentiation

Makoto Osanai, Jun Yamaguchi, Keisuke Kikuchi, Masaaki Satoh, Norimasa Sawada

Medical Electron Microscopy 37 ( 3 ) 193 - 197 2004年

We present a case of primitive neuroectodermal tumor (PNET) showing a unique differentiation phenotype based on ultrastructural observation. Rapidly growing tumor involving the retroperitoneum of a 68-year-old woman was characterized by histological findings including proliferation of diffuse poorly differentiated small round cells with scattered rosette formation similar to Homer-Wright type and by the ultrastructure demonstrating cytoplasmic neurosecretary granules and short cytoplasmic cilial structures. These cells revealed immunoreactivity only with neuron-specific enolase (NSE). We also observed a chromosomal translocation, t (11 22) (q24 q12), which is an identical recurrent alteration found in the neoplastic cells in the spectrum of PNETs. These findings may support the explanation that tumor cells observed in our case shared phenotypes of both neuronal and ependymal cell lineages and give a unique insight suggesting the possible histogenesis of PNETs. © The Clinical Electron Microscopy Society of Japan 2004.

Tight junctions and human diseases

Norimasa Sawada, Masaki Murata, Keisuke Kikuchi, Makoto Osanai, Hirotoshi Tobioka, Takashi Kojima, Hideki Chiba

Medical Electron Microscopy 36 ( 3 ) 147 - 156 2003年09月

書評論文，書評，文献紹介等

Tight junctions are intercellular junctions adjacent to the apical end of the lateral membrane surface. They have two functions, the barrier (or gate) function and the fence function. The barrier function of tight junctions regulates the passage of ions, water, and various macromolecules, even of cancer cells, through paracellular spaces. The barrier function is thus relevant to edema, jaundice, diarrhea, and blood-borne metastasis. On the other hand, the fence function maintains cell polarity. In other words, tight junctions work as a fence to prevent intermixing of molecules in the apical membrane with those in the lateral membrane. This function is deeply involved in cancer cell biology, in terms of loss of cell polarity. Of the proteins comprising tight junctions, integral membrane proteins occludin, claudins, and JAMs have been recently discovered. Of these molecules, claudins are exclusively responsible for the formation of tight-junction strands and are connected with the actin cytoskeleton mediated by ZO-1. Thus, both functions of tight junctions are dependent on the integrity of the actin cytoskeleton as well as ATP. Mutations in the claudin14 and the claudin16 genes result in hereditary deafness and hereditary hypomagnesemia, respectively. Some pathogenic bacteria and viruses target and affect the tight-junction function, leading to diseases. In this review, the relationship between tight junctions and human diseases is summarized.

Hepatocyte nuclear factor (HNF)-4 alpha triggers formation of functional tight junctions and establishment of polarized epithelial morphology in F9 embryonal carcinoma cells

H Chiba, T Gotoh, T Kojima, S Satohisa, K Kikuchi, M Osanai, N Sawada

EXPERIMENTAL CELL RESEARCH ( ACADEMIC PRESS INC ELSEVIER SCIENCE ) 286 ( 2 ) 288 - 297 2003年06月

F9 murine embryonal carcinoma cells provide an attractive system for facilitating molecular mechanisms for epithelial morphogenesis, since they have the capability of differentiating into polarized epithelial cells bearing an apical junctional complexes. We previously showed that a specific retinoid X receptor-retinoic acid receptor heterodimer transduced retinoid signals for biogenesis of functional tight junctions in F9 cells (Exp. Cell Res. 263, (2001) 163). In the present study we generated F9 cells expressing doxycycline-inducible hepatocyte nuclear factor (HNF)-4alpha, a nuclear receptor. We herein show that induction of HNF-4alpha initiates differentiation of F9 cells to polarized epithelial cells, in which tight-junction proteins occludin, claudin-6, claudin-7, and ZO-1 are concentrated at the apical-most regions of lateral membranes. Expression of occludin, claudin-6, and claudin-7 was induced in the cells by doxycycline treatment in a dose- and time-dependent manner, in terms of the amount of HNF-4a. In contrast, expression levels of ZO-1, ZO-2, E-cadherin, and P-catenin were not altered by HNF-4a. We also demonstrate, by analysis of diffusion of labeled sphingomyelin, that the fence function of tight junctions is achieved by induction of HNF-4a. These findings indicate that HNF-4a triggers de novo formation of functional tight junctions and establishment of epithelial cell polarity. (C) 2003 Elsevier Science (USA). All rights reserved.

Functions of hepatocyte nuclear factor-4 alpha in epithelial differentiation and proliferation of F9 embryonal carcinoma cells

H Chiba, T Goto, T Itoh, K Kikuchi, M Osanai, T Kojima, N Sawada

MOLECULAR BIOLOGY OF THE CELL ( AMER SOC CELL BIOLOGY ) 13 494A - 495A 2002年11月

研究発表ペーパー・要旨（国際会議）

Hepatocyte nuclear factor (HNF)-4 alpha induces expression of endothelial Fas ligand (FasL) to prevent cancer cell transmigration: A novel defense mechanism of endothelium against cancer metastasis

M Osanai, H Chiba, T Kojima, M Fujibe, K Kuwahara, H Kimura, M Satoh, N Sawada

JAPANESE JOURNAL OF CANCER RESEARCH ( BUSINESS CENTER ACADEMIC SOCIETIES JAPAN ) 93 ( 5 ) 532 - 541 2002年05月

Endothelial Fas ligand (FasL) contributes to the "immune privilege" of tissues such as testis and eye, in which apoptosis is induced in infiltrating Fas-positive activated T cells and results in the inhibition of leukocyte extravasation. In this study, we examined the role of endothelial FasL in controlling cancer cell transmigration using rat lung endothelial (RLE) cell line bearing a doxycycline-inducible hepatocyte nuclear factor (HNF)-4alpha expression system. We showed that a detectable level of FasL was expressed in RLE cells and that this expression was markedly up-regulated and well correlated to the degree of HNF-4alpha expression in a time-dependent manner. When various cancer cells were overlaid on an RLE monolayer sheet, we examined the ability of endothelial FasL to induce massive apoptosis in Fas-expressing cancer cells and found a causal link to inhibition of the transmigration. Finally we showed that FasL was expressed in capillaries of the rat brain by immunohistochemical staining, suggesting that FasL serves its functions not only in vitro, but also in vivo. These results raise the possibility that HNF-4alpha is involved in regulating cancer cell transmigration by modulating the Fas-FasL system.

Transient increase in telomerase activity of proliferating fibroblasts and endothelial cells in granulation tissue of the human skin

M Osanai, T Tamaki, M Yonekawa, A Kawamura, N Sawada

WOUND REPAIR AND REGENERATION ( BLACKWELL PUBLISHING INC ) 10 ( 1 ) 59 - 66 2002年01月

Although granulation tissue formation is an important step for second-intention wound healing, the molecular events underlying this process are still poorly understood, To investigate the role of telomerose in the formation of granulation tissue, we measured the activity of this enzyme and determined the expression and localization of human telomerase reverse transcriptase mRNA using human skin samples, Telomerase activity in the tip of the granulation tissue where fibroblasts actively proliferate was detected at a level 5,6 +/- 1.5 times higher than that at the edge of the tissue when using a polymerase chain reaction-based telomeric repeat amplification protocol assay coupled with enzyme-linked immunosorbent assay, This, together with the findings from semiquantitative reverse transcriptase-polymerase chain reaction and in situ hybridization of human telomerase reverse transcriptase, revealed that proliferating cell nuclear antigen-positive fibroblasts and endothelial cells in the progressing granulation tissue showed de novo activation of telomerase with high human telomerase reverse transcriptase mRNA expression, This condition may be a prolongation of cellular replicative capacity taking advantage of the positive regulatory dynamics of cell growth, We conclude that the regulation of telomerase activity may play an important role in granulation tissue formation in wound healing.

Hepatocyte nuclear factor (HNF)-4α induces expression of endothelial Fas ligand (FasL) to prevent cancer cell transmigration: A novel defense mechanism of endothelium against cancer metastasis.

Osanai M, Chiba H, Kojima T, Fujibe M, Satoh M, Sawada N

Jpn J Cancer Res. 93 ( 5 ) 532 - 541 2002年

Endothelial Fas ligand (FasL) contributes to the "immune privilege" of tissues such as testis and eye, in which apoptosis is induced in infiltrating Fas-positive activated T cells and results in the inhibition of leukocyte extravasation. In this study, we examined the role of endothelial FasL in controlling cancer cell transmigration using rat lung endothelial (RLE) cell line bearing a doxycycline-inducible hepatocyte nuclear factor (HNF)-4α expression system. We showed that a detectable level of FasL was expressed in RLE cells and that this expression was markedly up-regulated and well correlated to the degree of HNF-4α expression in a time-dependent manner. When various cancer cells were overlaid on an RLE monolayer sheet, we examined the ability of endothelial FasL to induce massive apoptosis in Fas-expressing cancer cells and found a causal link to inhibition of the transmigration. Finally, we showed that FasL was expressed in capillaries of the rat brain by immunohistochemical staining, suggesting that FasL serves its functions not only in vitro, but also in vivo. These results raise the possibility that HNF-4α is involved in regulating cancer cell transmigration by modulating the Fas-FasL system.

Transient increase in telomerase activity of proliferating fibroblasts and endothelial cells in granulation tissue of the human skin

Makoto Osanai, Tohru Tamaki, Motoki Yonekawa, Akio Kawamura, Norimasa Sawada

Wound Repair and Regeneration 10 ( 1 ) 59 - 66 2002年

Although granulation tissue formation is an important step for second-intention wound healing, the molecular events underlying this process are still poorly understood. To investigate the role of telomerase in the formation of granulation tissue, we measured the activity of this enzyme and determined the expression and localization of human telomerase reverse transcriptase mRNA using human skin samples. Telomerase activity in the tip of the granulation tissue where fibroblasts actively proliferate was detected at a level 5.6 ± 1.5 times higher than that at the edge of the tissue when using a polymerase chain reaction-based telomeric repeat amplification protocol assay coupled with enzyme-linked immunosorbent assay. This, together with the findings from semiquantitative reverse transcriptase-polymerase chain reaction and in situ hybridization of human telomerase reverse transcriptase, revealed that proliferating cell nuclear antigen-positive fibroblasts and endothelial cells in the progressing granulation tissue showed de novo activation of telomerase with high human telomerase reverse transcriptase mRNA expression. This condition may be a prolongation of cellular replicative capacity taking advantage of the positive regulatory dynamics of cell growth. We conclude that the regulation of telomerase activity may play an important role in granulation tissue formation in wound healing.

Unique cellular features in atypical adenomatous hyperplasia of the lung: Ultrastructural evidence of its cytodifferentiation

M Osanai, T Igarashi, Y Yoshida

ULTRASTRUCTURAL PATHOLOGY ( TAYLOR & FRANCIS INC ) 25 ( 5 ) 367 - 373 2001年09月

Atypical adenomatous hyperplasia (AAH) of the lung could be a good material to understand the histogenesis of peripherally occurring, well-differentiated adenocarcinoma. However, its true biological significance remains to be clarified. The authors present the histomorphological studies of this lesion and compare the ultrastructure with that of nonmucinous bronchioloalveolar carcinoma (BAC) to define characteristic features of AAH. Light microscopy showed the well-preserved pulmonary architecture, proliferated neoplastic cells without marked cellular atypia, and no transitional area to obvious adenocarcinoma. Intranuclear inclusion was present in a large number of neoplastic cells. Electron microscopy revealed that cuboidal or low columnar neoplastic cells proliferated actively but were not crowded on slightly thickened fibrous alveolar septa with both Clara-like granules and small lamellar bodies in the cytoplasm resembling that of Clara cell and type 2 pneumocyte. Some of the nuclei had characteristic invaginations of its nuclear membrane. Although the findings appear to be nonspecific for AAH, the authors emphasize that AAH is an alveolar intraepithelial neoplasia that represents a very early stage in the continuous developmental spectrum of adenomatous neoplasia in the bronchioloalveolar region corresponding to dysplasia or intraepithelial neoplasia in other organs, and will give the significance to speculate its histogenesis.

Adenocarcinoma arising in gastric heterotopic pancreas: Clinicopathological and immunohistochemical study with genetic analysis of a case

M Osanai, N Miyokawa, T Tamaki, M Yonekawa, A Kawamura, N Sawada

PATHOLOGY INTERNATIONAL ( BLACKWELL PUBLISHING ASIA ) 51 ( 7 ) 549 - 554 2001年07月

Heterotopic pancreas in the stomach is a relatively common congenital condition, but the risk of malignant transformation is extremely low. In this study, we describe a case of adenocarcinoma arising from a gastric heterotopic pancreas and we consider its morphological and immunohistochemical features and genetic analysis, in order to examine its histogenesis. This unusual sequela was seen in a 57-year-old woman. Image studies showed a protruding lesion with a central ulcer located in the lesser curvature from the angle to the body of the stomach. A biopsy specimen: confirmed this lesion as adenocarcinoma before total gastrectomy. The tumor showed mixed patterns of solid neoplastic-cell proliferation and moderately differentiated glandular structures, and also showed transitional lesions to obvious malignancy, that is, dysplasia, or adenocarcinoma in situ. Neoplastic cells had positive immunoreactivity for carbohydrate antigen (CA) 19-9. mucin (MUC) 1, and insulin, and the mutant allele-specific amplification method revealed a point mutation at K-ras codon 12 (GGT [Gly]-->GAT [Asp]), which is the most common mutational change observed in patients with pancreatic carcinoma. The features of the present case provide clear evidence that this tumor originated from heterotopic pancreatic tissue rather than from gastric epithelium.

Adenocarcinoma arising in gastric heterotopic pancreas: Clinicopathological and immunohistochemical study with genetic analysis of a case

M Osanai, N Miyokawa, T Tamaki, M Yonekawa, A Kawamura, N Sawada

PATHOLOGY INTERNATIONAL ( BLACKWELL PUBLISHING ASIA ) 51 ( 7 ) 549 - 554 2001年07月

Occludin and claudin-1 concentrate in the midbody of immortalized mouse hepatocytes during cell division

T Kojima, Y Kokai, H Chiba, M Osanai, K Kuwahara, M Mori, Y Mochizuki, N Sawada

JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY ( HISTOCHEMICAL SOC INC ) 49 ( 3 ) 333 - 339 2001年03月

It has been believed that epithelial cells maintain tight junctions at all times, including during cell division, to provide a continuous epithelial seal. However, changes in localization of integral tight junction proteins during cell division have not been examined. In this study, using SV40-immortalized mouse hepatocytes transfected with human Cx32 cDNA, in which tight junction strands and the endogenous tight junction proteins occludin, claudin-1, ZO-1, and ZO-2 were induced, we examined changes in localization of the tight junction proteins at all stages of cell division. All tight junction proteins were present between mitotic cells and neighboring cells throughout cell division. in late telophase, the integral tight junction proteins occludin and claudin-1, but not the cytoplasmic proteins ZO-1 and ZO-2, were concentrated in the midbody between the daughter cells and were observed at cell borders between the daugher and neighboring cells. These results indicate that the integral tight junction proteins are regulated in a different manner from the cytoplasmic proteins ZO-1 and ZO-2 during cytokinesis.

Retinoid X receptor alpha and retinoic acid receptor gamma mediate expression of genes encoding tight-junction proteins and barrier function in F9 cells during visceral endodermal differentiation

H Kubota, H Chiba, Y Takakuwa, M Osanai, H Tobioka, GI Kohama, M Mori, N Sawada

EXPERIMENTAL CELL RESEARCH ( ACADEMIC PRESS INC ) 263 ( 1 ) 163 - 172 2001年02月

Retinoids are critical for differentiation of columnar epithelial cells and for preventing metaplasia of these cells into stratified squamous epithelial cells, in which tight junctions (TJs) are essentially absent. This implies that retinoids might play important roles in regulating the structures and functions of TJs of columnar epithelium. F9 murine embryonal carcinoma cells differentiate into epithelial cells resembling visceral endoderm bearing TJs, when grown in suspension as aggregates in the presence of retinoic acid (RA). We show that RA induces the TJ structure and expression of several TJ-associated molecules, such as ZO-1, occludin, claudin-6, and claudin-7, as well as a barrier function in the genetically engineered cell line F9: rtTA:Cre-ERT L32T2, which allows sophisticated genetic manipulations simply by addition of ligands (H. Chiba et at, 2000, Exp. Cell Res. 260, 334-339). Interestingly, our data indicate that a barrier for small substances is generated after that for large ones during de novo formation of TJs, We also compared the RA-induced expression of TJ components and barrier function in RXR alpha (-/-)-RAR gamma (-/-) F9 cells with those in wild-type cells and show that the retinoid signals for transduction of these events are mediated by specific RXR-RAR pairs. (C) 2001 Academic Press.

Expression of GFR alpha-1, receptor for GDNF, in rat brain capillary during postnatal development of the BBB

H Utsumi, H Chiba, Y Kamimura, M Osanai, Y Igarashi, H Tobioka, M Mori, N Sawada

AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY ( AMER PHYSIOLOGICAL SOC ) 279 ( 2 ) C361 - C368 2000年08月

It is well known that the blood-brain barrier (BBB) matures at similar to 2 wk after birth in the rat. Recently, we showed that glial cell line-derived neurotrophic factor (GDNF) enhances the barrier function of porcine endothelial cells forming the BBB in culture. In the present study, we examined the relation between permeability of the BBB, using Evans blue as a tracer, and expression of the GDNF family receptor (GFR alpha-1) during postnatal development of the BBB. Morphometric analysis showed that exudation of Evans blue from capillaries of the cerebral cortex progressively decreased until postnatal day 21. Inversely, immunohistochemical examinations showed expression of GFR alpha-1 in the capillaries at postnatal day 3 and expression that reached the same levels as observed in adult rats by postnatal day 10. However, c-ret, which is thought to mediate a signal evoked by binding of GDNF to GFR alpha-1, was not expressed in the capillaries of the brain cortex in 3-mo-old rats. On the other hand, the tight junction proteins occludin and ZO-1 appeared to be fully expressed at birth. The reciprocal relation between GFR alpha-1 expression and the permeability of the BBB strongly suggests active participation of GDNF in postnatal development of the BBB, although the mechanism(s) involved is still veiled.

Gain of chromosomes 15 and 19 is frequent in both mouse hepatocellular carcinoma cell lines and primary tumors, but loss of chromosomes 4 and 12 is detected only in the cell lines

K Ogawa, M Osanai, M Obata, K Ishizaki, K Kamiya

CARCINOGENESIS ( OXFORD UNIV PRESS ) 20 ( 11 ) 2083 - 2088 1999年11月

Chromosomal alterations were investigated in hepatocellular carcinoma cell lines, primary tumors and liver epithelial cell lines derived from normal livers of C57BL/6J x C3H/HeJ F-1 and C3H/HeJ x C57BL/6J F-1 mice. In the primary tumors, non-random gain of chromosomes 15 and 19 was found in seven and five of 14 hepatocellular carcinomas, respectively. On the other hand, in the cases of both liver epithelial and hepatocellular carcinoma cell lines, frequent changes were loss of chromosomes 4 (4/9 cell lines) and 12 (3/9) as well as gain of chromosomes 15 (5/9) and 19 (4/9), These results indicate that the chromosomal gain is associated with both in vivo carcinogenesis and establishment of cell lines, while the loss is specific for the latter. PCR analysis using polymorphic microsatellite DNA markers revealed that the loss of chromosome 12 as well as chromosome 4 was much more frequent for the C57BL/6J hepatocarcinogenesis-resistant rather than the susceptible C3H/HeJ strain.

Morphology, proliferation and apoptosis of mouse liver epithelial cells cultured as spheroids

GH Lee, M Osanai, Y Tokusashi

JAPANESE JOURNAL OF CANCER RESEARCH ( ELSEVIER SCI IRELAND LTD ) 90 ( 10 ) 1109 - 1116 1999年10月

The MLEC10 is an epithelial cell line derived from an untreated, normal C3H/HeN mouse liver We previously demonstrated that tumorigenic variants from this cell line produced moderately differentiated hepatocellular carcinomas in nude mice, However, it has remained unclear whether the parental MLEC10 cells represent immortalized hepatocytes or so-called oval cells, both of which may serve as precursors for hepatocellular neoplasms. In this study, we performed 3-dimensional, spheroid culture of the MLEC10 cells in order to facilitate histological assessment of their lineage. Spheroidal aggregates were formalin-fixed and embedded in paraffin for routine light-microscopic observation of hematoxylin and eosin-stained sections. Histopathologically, the MLEC10 cells were indistinguishable from immature hepatocytes and distinct from oval cells. At the electron-microscopic level, their hepatocytic nature was evidenced by bile canaliculus structures and glycogen storage. Intriguingly, the spheroids contained fragmentary material reminiscent of Councilman bodies, implying apoptosis of the hepatocytes. Although the cells significantly proliferated during the first three days of culture, apoptotic death then resulted in a 75% decrease in viable cell number. Thereafter, both apoptosis and cell division appeared silent, the numbers being unchanged. Expression of the p53 tumor suppressor gene became gradually elevated, correlating positively with growth arrest, but negatively with apoptosis, suggesting that the cell death occurred independently of p53, Our results indicate that at least some liver epithelial cell lines derived from untreated murine livers exhibit a hepatocytic morphology in spheroid culture. Also, the present culture system provides a useful tool for investigating biological phenomena, e.g. apoptosis, specifically involving liver cells, under 3-dimensional conditions.

Cryofiltration: A new approach for removal of anti-blood group antibody

M Takahashi, T Tamaki, M Tanaka, M Katori, N Yokota, Y Takamine, M Osanai, M Naitoh, M Yasuhara, K Kukita, J Meguro, M Yonekawa, A Kawamura

TRANSPLANTATION PROCEEDINGS ( ELSEVIER SCIENCE INC ) 30 ( 7 ) 3275 - 3276 1998年11月

Mechanism of the paradoxical, inhibitory effect of phenobarbital on hepatocarcinogenesis initiated in infant B6C3F(1) mice with diethylnitrosamine

GH Lee, T Ooasa, M Osanai

CANCER RESEARCH ( AMER ASSOC CANCER RESEARCH ) 58 ( 8 ) 1665 - 1669 1998年04月

Phenobarbital (PB), a classical rodent hepatopromoter, remarkably enhances hepatocarcinogenesis initiated by diethylnitrosamine (DEN) in adult B6C3F(1), mice. However, it is also known to strongly inhibit liver tumor development in the B6C3F(1) mice initiated with DEN in their infancy. The present study aimed to elucidate the unknown biological mechanisms for this paradoxical, inhibitory effect of PB on B6C3F(1) mouse hepatocarcinogenesis. Male 12-day-old infant B6C3F(1) mice were injected i.p. with DEN and, at 6 weeks of age, divided into PB-treated (PB+ group) and untreated (PB-group) animals. At 24 weeks, PB treatment was ceased for half of the PB+ animals (PB+/- group) and started for half of the PB-animals (PB-/+ group), Finally, all mice were sacrificed at 36 weeks and examined for the development of liver tumors. The mean multiplicity of gross tumors in the PB+ group was only one-fifteenth of that for the PB-group. PB-/+ animals developed fewer than half of the tumors found in PB-mice, indicating that the PB effect depends solely on the treatment duration, rather than the animal age, The effect was proven to be reversible, because the mean tumor multiplicity for the PB+/- group was seven times larger than that for the PB+ group. Stereological analysis revealed the mean volume of hepatocellular proliferative lesions in the PB-animals to be 7.7- and 4.1-fold the values for the PB+ and PB-/+ groups, respectively. The mean proliferating cell nuclear antigen-labeling indices for hepatocellular adenomas in PB+ and PB-/+ animals were also one-third of that for tumors in PB-animals, whereas no significant differences were observed with regard to the mean apoptotic index. In conclusion the inhibitory effect of PB seemed to be primarily caused by the suppression of tumor tell proliferation. Irrespective of the group, most lesions observed were basophilic hepatocellular adenomas or foci, positive for Bcl-2 oncoprotein, They were thus distinct from the eosinophilic Bcl-2- lesions that predominate with PB promotion after the initiation of adult B6C3F(1) mice. This age-dependent nature of initiation, together with the differential responses of Bcl-2+ and Bcl-2- lesions, may be responsible for the apparently contradictory outcomes of PB treatment in infant and adult B6C3F(1) mice.

Cryofiltration apheresis for major ABO-incompatible kidney transplantation

Tohru Tamaki, Mitsuko Tanaka, Masamichi Katori, Makoto Osanai, Mitsuo Yasuhara, Jun-Ichi Meguro, Kazutaka Kukita, Motoki Yonekawa, Akio Kawamura

Therapeutic Apheresis ( Blackwell Publishing Inc. ) 2 ( 4 ) 308 - 310 1998年

The barrier of ABO-incompatible kidney transplantation is the presence of anti-A and anti-B antibodies in the recipient's circulating blood. Double filtration plasmapheresis (DFPP) is usually used to eliminate those antibodies. We tried cryofiltration apheresis (CRYO) in 2 recipients. Patient 1 was a 45-year-old male with B, Rh(+). The titers of IgM anti-A antibody were only reduced from x64 to x32 by the end of 3 sessions of standard CRYO. Renal allografting was not performed. Case 2 was a 29-year-old male with B, Rh(+). CRYO was introduced for 3 sessions. The initial IgM and IgG titers were x128 and negative, respectively. The standard CRYO system was modified by temperature, treated volume, and filter pore size. The IgM anti-A antibody titer was markedly reduced to x2 after the final session of CRYO. The donor was a 56-year-old father with A, Rh(+). Tacrolimus, azathioprine, methylprednisolone, and antilymphocyte globulin were used as the introductory immunosuppression therapy.

Immunomodulation in transplant patients by cryofiltration

Akio Kawamura, Makoto Osanai, Motoki Yonekawa

Therapeutic Apheresis ( Blackwell Publishing Inc. ) 2 ( 3 ) 205 - 209 1998年

書評論文，書評，文献紹介等

Cryofiltration was applied to major ABO incompatible bone marrow transplant (BMT), renal transplant, and chronic renal rejection cases. The anti-ABO titers IgM and IgG pretreatment by cryofiltration were x64 and x8 in a major ABO incompatible BMT patient. After treatments, they were decreased to x16 and x2, respectively. BMT was performed successfully. In a case of ABO major incompatible renal transplant, anti-A-IgM before cryofiltration treatment was x128, and CH50 was 26.5. Three treatments were performed. As a result, the anti-A-IgM and CH50 levels were decreased to x4 and under 10. After renal transplantation, antibodies appeared again, but because the creatinine level was normal, cryofiltration was not performed again. Five cases of chronic rejection of renal transplants were treated by cryofiltration. They remained hemodialysis (HD) free for an average of 17.4 months, and the longest duration was 32 months. Cryofiltration is a good treatment for extracorporeal immunomodulation in transplantation.

Basic clinical study of an easy and effective leukocytapheresis by the use of nonwoven polyester filter

Akio Kawamura, Makoto Osanai, Masamichi Katori, Motoki Yonekawa, Masao Saitoh

Therapeutic Apheresis ( Blackwell Publishing Inc. ) 2 ( 4 ) 292 - 296 1998年

Leukocytapheresis (LCAP) is widely used for the treatment of immunological diseases. We studied a new treatment of LCAP using a nonwoven polyester filter. In a basic study, 30-70% of the lymphocytes were adsorbed. Also, 30-68% of the lymphocyte subsets were removed. This method was applied to 2 patients with corticosteroid-resistant active ulerative colitis. Erosion, edema, bleeding, ulcer formation, and stenosis of the colon were almost completely repaired after 6 LCAP treatments. LCAP using a nonwoven polyester filter will be a very useful treatment for immunological diseases and extracorporeal immunomodulation.