精神疾患に関与するポリシアル酸転移酵素の機能解析

• Project Area: Deciphering sugar chain-based signals regulating integrative neuronal functions
• Project/Area Number: 26110708
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Nagoya University
• Principal Investigator: 佐藤 ちひろ 名古屋大学, 生物機能開発利用研究センター, 准教授 (10343211)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥9,360,000 (Direct Cost: ¥7,200,000、Indirect Cost: ¥2,160,000); Fiscal Year 2015: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2014: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
• Keywords: ポリシアル酸 / 統合失調症 / BDNF / 分子間相互作用 / 糖鎖 / ポリシアル酸転移酵素 / 精神疾患 / 自閉症 / 気分障害

Outline of Annual Research Achievements

本研究は統合失調症などの精神疾患に関連するポリシアル酸転移酵素ST8SIA2の機能解析が目的であり、本研究では報告のある精神疾患患者に有意なSNPsや変異に関して、それらの酵素がもたらす産物であるポリシアル酸の構造および機能解析を行った。具体的にはプロモーターに存在するrSNPに関してはプロモータ解析および遺伝子のマイクロアレイの解析を各種細胞を用いて行い、ある転写因子によって細胞特異的にST8SIA2の発現異常が起こる可能性を示した。翻訳領域に存在しアミノ酸置換を伴うSNPの解析から、疾患型が合成するポリシアル酸が損なわれ、特にBDNFの結合やproBDNFの結合性が正常型より損なわれることがわかった。また翻訳領域に存在しアミノ酸置換を伴わないsSNPは、コドン使用頻度によりST8SIA2の翻訳速度が変化し、ポリシアル酸の生合成に影響を与える可能性が示唆された。イントロンに存在するiSNPは、特に双極性障害型のSNPに関して、マウスの神経細胞およびヒトHEK細胞においてST8SIA2の合成が促進され、polySiaが過剰発現することが明らかになった。ST8SIA2の過剰発現の影響を調べるために子宮内エレクトロポレーション法により、ST8SIA2およびST8SIA4の遺伝子導入を行ったところ、ニューロンの移動がMockに比べて変化した。以上のことより、ST8SIA2遺伝子発現は細胞特異的に発現が厳密に制御されており、それに伴いpolySiaの構造が損なわれることが分かった。つまり、polySia構造は厳密に構造が制御されており、それによって神経作用因子であるBDNF、FGF2, proBDNFの結合性を制御することなどで、正常な神経機能を果たすことが考えられた。

Research Progress Status

27年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

27年度が最終年度であるため、記入しない。

Research Products

• [Int'l Joint Research] UC San Diego(米国)
• [Int'l Joint Research]
• [Journal Article] Rapid trimming of cell surface polySia by exovesicular sialidase triggers release of preexisting surface neurotrophin. (2015)
  • Author(s): Sumida M, Hane M, Yabe U, Shimoda Y, Pearce OM, Kiso M, Miyagi T, Sawada M, Varki A, Kitajima K, and Sato C.
  • Journal Title: J. Biol. Chem. Volume: 290 Pages: 13202-14
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Protective effects of polysialic acid on proteolytic cleavage of FGF2 and proBDNF/BDNF. (2015)
  • Author(s): Hane M, Matsuoka S, Miyata S. Kitajima K and Sato C.
  • Journal Title: Glycobiology Volume: 10 Pages: 1112-24
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Advanced technologies in sialic acid and sialoglycoconjugate anaysis. (2015)
  • Author(s): Kitajima K, Varki N, Sato C.
  • Journal Title: Top Curr Chem. Volume: 367 Pages: 75-103
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Sulfatide-Hsp70 interaction promotes Hsp70 clustering and stabilizes binding to unfolded protein. (2015)
  • Author(s): Harada Y, Sato C, Kitajima K.
  • Journal Title: Biomolecules Volume: 15 Pages: 958-73
  • Peer Reviewed
• [Journal Article] Discovery, primary and crystal structures, and capacitation-related properties of a prostate-derived heparin-binding protein WGA16 from boar sperm. (2015)
  • Author(s): Garénaux E, Kanagawa M, Tsuchiyama T, Hori K, Kanazawa T, Goshima A, Chiba M, Yasue H, Ikeda A, Yamaguchi Y, Sato C, and Kitajima K.
  • Journal Title: J. Biol. Chem. Volume: 290 Pages: 5484-5501
  • Peer Reviewed
• [Journal Article] Ganglioside contained in the neuronal tissue-enriched acidic protein of 22 kDa (NAP-22) fraction prepared from the detergent-resistant membrane microdomain of rat brain inhibits the phosphatase activity of calcineurin. (2015)
  • Author(s): Kobayashi Y, da Silva R, Kumanogoh H, Miyata S, Sato C, Kitajima K, Nakamura S, Morita M, Hayashi F, Maekawa S.
  • Journal Title: .J Neurosci Res. Volume: 93 Pages: 1462-70
  • Peer Reviewed
• [Journal Article] Polysialic acid: Biosynthesis, novel functions and applications. Critical Reviews in Biochemistry and Molecular Biology (2014)
  • Author(s): Karen Colley, Ken Kitajima, Chihiro Sato
  • Journal Title: Critical Reviews in Biochemistry and Molecular Biology Volume: - Issue: 6 Pages: 1-35
  • DOI: 10.3109/10409238.2014.976606
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Comparison of analytical methods to detect polysialic acid. (2014)
  • Author(s): Saki Nishimura, Masaya Hane, Yuki Niimi, Shinji Miyata, Ken Kitajima, Chihiro Sato
  • Journal Title: Journal of Glycomics and Lipidomics Volume: 4 Pages: 113-113
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] ST8 alpha-N-acetyl-neuraminide alpha-2,8-sialyltransferase 2 (ST8SIA2) (2014)
  • Author(s): Chihiro Sato
  • Journal Title: Handbook of Glycosyltransferases and related genes Volume: - Pages: 781-795
  • Peer Reviewed
• [Journal Article] Frontal Affinity Chromatography in the Methods in Molecular Biology (2014)
  • Author(s): Chihiro Sato
  • Journal Title: Lectins Volume: - Pages: 257-264
  • Peer Reviewed
• [Journal Article] Polysialic acid (2014)
  • Author(s): Chihiro Sato and Ken Kitajima
  • Journal Title: Glycoscience: Biology and Medicine Volume: - Pages: 519-528
  • Peer Reviewed
• [Journal Article] Polysialyltransferase assay (2014)
  • Author(s): Masaya Hane, Ken Kitajima and Chihiro Sato
  • Journal Title: Glycoscience: Biology and Medicine Volume: - Pages: 529-534
  • Peer Reviewed
• [Journal Article] Flagellasialin (2014)
  • Author(s): Takeru Kanazawa, Eriko Suzuki, Shinji Miyata, Chihiro Sato, and Ken Kitajima
  • Journal Title: Glycoscience: Biology and Medicine Volume: - Pages: 883-890
  • Peer Reviewed
• [Journal Article] Interaction of 70-kDa heat shock protein with glycosaminoglycans and acidc glycopolymers. (2014)
  • Author(s): Harada Yoichiro, Estelle Garenaux, Takehiro Nagatsuka, Hirotaka Uzawa, Yoshihiro Nishida, Chihiro Sato, and Ken Kitajima
  • Journal Title: Biochem Biophys Res Commun Volume: 453 Pages: 229-234
  • Peer Reviewed
• [Journal Article] Tissue-specific post-translational modification allows functional targeting of thyrotropin. (2014)
  • Author(s): Keisuke Ikegami, Xia-Hui Liao, Yuta Hoshino, Hiroko Ono, Wataru Ota, Yuka Ito, Taeko Nishiwaki-Ohkawa, Chihiro Sato, Ken Kitajima, Masayuki Iigo, Yasufumi Shigeyoshi, Masanobu Yamada, Yoshiharu Murata, Samuel Refetoff, Takashi Yoshimura.
  • Journal Title: Cell Reports Volume: 9 Pages: 1-9
  • Peer Reviewed
• [Presentation] Effects of the genetic alterations of ST8SIA2/STX reported from psychiatric diseases on the structure and functions of polysialic acid. (2016)
  • Author(s): Chihiro Sato.
  • Organizer: 3rd International symposium on Glyco-neuroscience
  • Place of Presentation: Hyogo, Japan
  • Year and Date: 2016-01-14
  • Int'l Joint Research / Invited
• [Presentation] Brain-region specific change of polysialic acid (polySia) on NCAM under acute stress conditions. (2016)
  • Author(s): Chikara Abe, Masaya Hane, Sinji Miyata, Ken Kitajima, and Chihiro Sato.
  • Organizer: 3rd International symposium on Glyco-neuroscience
  • Place of Presentation: Hyogo, Japan
  • Year and Date: 2016-01-14
  • Int'l Joint Research
• [Presentation] ポリシアル酸転移酵素STX/ST8SIA2とPST/ST8SIA4は異なるポリシアル酸構造および神経作用因子結合性を生み出す (2015)
  • Author(s): 新美百希、羽根正弥、北島健、佐藤ちひろ
  • Organizer: 第88回日本生化学会大会・第38回日本分子生物学会年会合同大会
  • Place of Presentation: 神戸
  • Year and Date: 2015-12-01
  • Int'l Joint Research
• [Presentation] 高次脳機能や精神疾患に関与するポリシアル酸構造の新機能 (2015)
  • Author(s): 佐藤ちひろ
  • Organizer: 日本薬学会東海支部特別講演会
  • Place of Presentation: 静岡県立大学
  • Year and Date: 2015-10-23
  • Invited
• [Presentation] Structural and functional changes of polysialic acid related with genetic alterations of ST8SIA2/STX in psychiatric diseases. (2015)
  • Author(s): Chihiro Sato.
  • Organizer: 23rd International Symposium on Glycoconjugates (Glyco23)
  • Place of Presentation: 23rd International Symposium on Glycoconjugates (Glyco23)
  • Year and Date: 2015-09-15
  • Int'l Joint Research
• [Presentation] Effects of psychiatric disorders-related intronic single nucleotide polymorphisms (iSNPs) of the polysialyltransferase ST8SIA2/STX gene. (2015)
  • Author(s): Masaya Hane, Ken Kitajima, Chihiro Sato.
  • Organizer: 23rd International Symposium on Glycoconjugates (Glyco23)
  • Place of Presentation: 23rd International Symposium on Glycoconjugates (Glyco23)
  • Year and Date: 2015-09-15
  • Int'l Joint Research
• [Presentation] ポリシアル酸転移酵素STX/ST8SIA2とPST/ST8SIA4のポリシアル酸生合成能の解析 (2015)
  • Author(s): 新美百希, 羽根正弥, 松岡修平, 宮田真路, 北島健, 佐藤ちひろ
  • Organizer: 第34回日本糖質学会年会
  • Place of Presentation: 東京
  • Year and Date: 2015-07-31
• [Presentation] マウスの急性ストレス条件下における脳内ポリシアル酸構造の領域特異的変動の解析. (2015)
  • Author(s): 阿部智佳羅, 羽根正弥, 宮田真路, 北島健, 佐藤ちひろ
  • Organizer: 第34回日本糖質学会年会
  • Place of Presentation: 東京
  • Year and Date: 2015-07-31
• [Presentation] 哺乳類細胞の内在性ポリシアル酸転移酵素の性質の解析 (2015)
  • Author(s): 石原幸奈, 羽根正弥, 宮田真路, 北島 健、佐藤ちひろ
  • Organizer: 第34回日本糖質学会年会
  • Place of Presentation: 東京
  • Year and Date: 2015-07-31
• [Presentation] α2,6-シアル酸残基のメダカ初期胚における重要性. (2015)
  • Author(s): 36.北島健, 呉迪, 田嶋克枝, 丸山恵実, 足立朋子, 橋本寿史, 日比正彦, 藤田明子, 佐藤ちひろ.
  • Organizer: 日本農芸化学会2015年度岡山大会
  • Place of Presentation: 岡山
  • Year and Date: 2015-03-26 – 2015-03-29
• [Presentation] Comparison of enzyme properties of three insect CMP-sialic acid synthetases. (2015)
  • Author(s): Di Wu, Akiko Fujita, Chihiro Sato, Ken Kitajima.
  • Organizer: 日本農芸化学会2015年度岡山大会
  • Place of Presentation: 岡山
  • Year and Date: 2015-03-26 – 2015-03-29
• [Presentation] Insight into single nucleotide polymorphisms (SNPs) of the polysialyltransferase ST8SIA2/STX in psychiatric disorders. (2014)
  • Author(s): Masaya Hane, Saki Nishimura,Toshiyuki Hayakawa,Ken Kitajima, Chihiro Sato.
  • Organizer: 2014 SFG & JSCR Joint Meeting Satellite Symposium II Glycans in Neuroscience
  • Place of Presentation: Hawaii, USA
  • Year and Date: 2014-11-16 – 2014-11-19
• [Presentation] An anti-schizophrenic drug affects the surface expression of polySia-NCAM in IMR-32 human neuroblastoma cells. (2014)
  • Author(s): Saki Nishimura, Masaya Hane, Yuki Niimi, Ken Kitajima, Chihiro Sato.
  • Organizer: 2014 SFG & JSCR Joint Meeting
  • Place of Presentation: Hawaii, USA
  • Year and Date: 2014-11-16 – 2014-11-19
• [Presentation] Critical importance of the surface expression of α2,6-sialosides during early development of medaka fish. (2014)
  • Author(s): 34.Ken Kitajima, Di Wu, Katsue Tajima, Emi Maruyama, Yuko Yasukawa, Lan-Yi Chang, Yoshihito Taniguchi, Yasuhiro Kamei, Tomoko Adachi, Hisashi Hashimoto, Masahiko Hibi, Akiko Fujita, Chihiro Sato
  • Organizer: 2014 SFG & JSCR Joint Meeting
  • Place of Presentation: Hawaii, USA
  • Year and Date: 2014-11-16
• [Presentation] 昆虫CMP-シアル酸合成酵素の新奇な性質の発見． (2014)
  • Author(s): 呉迪、藤田明子、佐藤ちひろ、北島健．
  • Organizer: 第87回日本生化学会大会
  • Place of Presentation: 京都
  • Year and Date: 2014-10-15 – 2014-10-18
• [Presentation] シアル酸アルドラーゼSPLの基質特異性に関わるアミノ酸残基の同定． (2014)
  • Author(s): 熊澤慎介、藤田明子、佐藤ちひろ、北島健．
  • Organizer: 第87回日本生化学会大会
  • Place of Presentation: 京都
  • Year and Date: 2014-10-15 – 2014-10-18
• [Presentation] ポリシアル酸はNCAMの細胞外ドメイン切断を制御する機能を持つ． (2014)
  • Author(s): 松岡修平、羽根正弥、西村紗希、北島健、佐藤ちひろ
  • Organizer: 第87回日本生化学会大会
  • Place of Presentation: 京都
  • Year and Date: 2014-10-15 – 2014-10-18
• [Presentation] ポリシアル酸転移酵素ST8SIA2/STXのアミノ酸変異を伴わない一塩基多型変異(SNP)によるポリシアル酸生合成の制御. (2014)
  • Author(s): 羽根正弥、西村紗希、早川敏之、北島健、佐藤ちひろ.
  • Organizer: 第87回日本生化学会大会
  • Place of Presentation: 京都
  • Year and Date: 2014-10-15 – 2014-10-18
• [Presentation] ポリシアル酸がつくる細胞外環境とその生物学的意義. (2014)
  • Author(s): 佐藤ちひろ.
  • Organizer: 第87回日本生化学会大会
  • Place of Presentation: 京都
  • Year and Date: 2014-10-15 – 2014-10-18
  • Invited
• [Presentation] ヒト系統におけるシアル酸転移酵素ST8SiaIIの発現多型の進化． (2014)
  • Author(s): 早川敏之、藤戸尚子、羽根正弥、北島健、佐藤ちひろ、颯田葉子.
  • Organizer: 日本遺伝学会第86回大会
  • Place of Presentation: 滋賀
  • Year and Date: 2014-09-17 – 2014-09-19
• [Presentation] マウス雄性生殖器官におけるCD52の局在解析． (2014)
  • Author(s): 鈴木英里子、宮田真路、金澤尊、佐藤ちひろ、北島健.
  • Organizer: 日本動物学会第85回仙台大会
  • Place of Presentation: 仙台
  • Year and Date: 2014-09-11 – 2014-09-13
• [Presentation] Polysialic acid as an attractive molecule. (2014)
  • Author(s): Chihiro Sato, Ken Kitajima.
  • Organizer: Sialoglyco2014
  • Place of Presentation: Gold Coast, Australia
  • Year and Date: 2014-09-07 – 2014-09-10
  • Invited
• [Presentation] Critical importance of the surface expression of α2,6-sialosides during early development of medaka, Oryzias latipes. (2014)
  • Author(s): Katsue Tajima, Emi Maruyama, Lan-Yi Chang, Adachi Tomoko, Hisashi Hashimoto, Masahiko Hibi, Chihiro Sato, Ken Kitajima.
  • Organizer: Sialoglyco2014
  • Place of Presentation: Gold Coast, Australia
  • Year and Date: 2014-09-07 – 2014-09-10
  • Invited
• [Presentation] Abnormality of the unique CMP-Sia synthetase-mutant medaka fishes established by the TILLING method. (2014)
  • Author(s): Di Wu, Akiko Fujita, Yuko Yasukawa, Yoshihito Taniguchi, Yasuhiro Kamei, Chihiro Sato, Ken Kitajima.
  • Organizer: Sialoglyco2014
  • Place of Presentation: Gold Coast, Australia
  • Year and Date: 2014-09-07 – 2014-09-10
• [Presentation] Analysis of a ligand-binding mechanism among sialic-acid binding sites in Siglec-7. Sialoglyco 2014; September 7-10, 2014; Crowne Plaza Surfers Paradise, Gold Coast, Australia (2014)
  • Author(s): Ami Goshima, Nao Yamakawa, Yu Yasuda, Hiroshi Tanaka, Paul Crocker, Ken Kitajima, Chihiro Sato.
  • Organizer: Sialoglyco2014
  • Place of Presentation: Gold Coast, Australia
  • Year and Date: 2014-09-07 – 2014-09-10
• [Presentation] Analysis of a ligand-binding mechanism among sialic-acid binding sites in Siglec-7. (2014)
  • Author(s): Saki Nishimura, Yuki Niimi, Masaya Hane, Ken Kitajima, Chihiro Sato.
  • Organizer: Sialoglyco2014
  • Place of Presentation: Gold Coast, Australia
  • Year and Date: 2014-09-07 – 2014-09-10
• [Presentation] Effects of Single Nucleotide Polymorphism (SNP)s in a polysialyltransferase ST8SIA2/STX found in psychiatric disorder patients. (2014)
  • Author(s): Masaya Hane, Saki Nishimura, Toshiyuki Hayakawa, Ken Kitajima, Chihiro Sato.
  • Organizer: Sialoglyco2014
  • Place of Presentation: Gold Coast, Australia
  • Year and Date: 2014-09-07 – 2014-09-10
• [Presentation] シアル酸転移酵素STXのプロモーター多型の進化と環境適応． (2014)
  • Author(s): 藤戸尚子、早川敏之、羽根正弥、北島健、佐藤ちひろ、颯田葉子．
  • Organizer: 日本進化学会第16回大阪大会
  • Place of Presentation: 大阪
  • Year and Date: 2014-08-21 – 2014-08-24
• [Presentation] メダカ初期発生にはシアル酸が必須である． (2014)
  • Author(s): 呉 迪、田嶋克枝、川本英里、丸山恵美、安川裕子、Chang Lan-Yi、谷口善仁、亀井保博、足立朋子、橋本寿史、日比正彦、藤田明子、佐藤ちひろ、北島 健．
  • Organizer: 第33回日本糖質学会年会
  • Place of Presentation: 名古屋
  • Year and Date: 2014-08-10 – 2014-08-12
• [Presentation] シアル酸は初期発生に必須である：TILLING法によるCMP-シアル酸合成酵素変異体メダカの樹立． (2014)
  • Author(s): 9.呉 迪､藤田明子、安川裕子、谷口善仁、亀井保博、佐藤ちひろ、北島 健．
  • Organizer: 第33回日本糖質学会年会
  • Place of Presentation: 名古屋
  • Year and Date: 2014-08-10 – 2014-08-12
• [Presentation] ポリシアル酸はNCAM細胞外ドメインの切断を制御する機能を持つ． (2014)
  • Author(s): 松岡修平、羽根正弥、新美百希、北島 健、佐藤ちひろ．
  • Organizer: 第33回日本糖質学会年会
  • Place of Presentation: 名古屋
  • Year and Date: 2014-08-10 – 2014-08-12
• [Presentation] Siglec-7 V-setドメインに存在する複数のシアル酸結合部位のリガンド結合制御の解析． (2014)
  • Author(s): 五島亜実、山川奈緒、安田優、田中浩士、宮田真路、北島健、佐藤ちひろ.
  • Organizer: 第33回日本糖質学会年会
  • Place of Presentation: 名古屋
  • Year and Date: 2014-08-10 – 2014-08-12
• [Presentation] 動物精子ラフトにおける糖鎖に富むペプチドの普遍的存在の証明 (2014)
  • Author(s): 鈴木英里子、宮田真路、岩尾康宏、佐藤ちひろ、北島健.
  • Organizer: 第33回日本糖質学会年会
  • Place of Presentation: 名古屋
  • Year and Date: 2014-08-10 – 2014-08-12
• [Presentation] ポリシアル酸転移酵素STX/ST8SIA2とPST/ST8SIA4のポリシアル酸生合成能の解析． (2014)
  • Author(s): 新美百希、羽根正弥、西村紗希、宮田真路、北島健、佐藤ちひろ.
  • Organizer: 第33回日本糖質学会年会
  • Place of Presentation: 名古屋
  • Year and Date: 2014-08-10 – 2014-08-12
• [Presentation] Siglec-7に存在する複数のシアル酸結合部位のリガンド結合制御機構の解析． (2014)
  • Author(s): 五島亜実、安田優、宮田真路、北島健、佐藤ちひろ
  • Organizer: 第78回日本生化学中部支部例会・シンポジウム
  • Place of Presentation: 名古屋
  • Year and Date: 2014-05-24
• [Presentation] 動物精子ラフトに局在する糖鎖に富むタンパク質の普遍的存在の証明． (2014)
  • Author(s): 新美百希、羽根正弥、松岡修平、3.新美百希、羽根正弥、松岡修平、北島健、佐藤ちひろ、宮田真路、北島健、佐藤ちひろ
  • Organizer: 第78回日本生化学中部支部例会・シンポジウム
  • Place of Presentation: 名古屋
  • Year and Date: 2014-05-24
• [Presentation] 神経活動を司るポリシアル酸転移酵素STXとPSTの機能解析． (2014)
  • Author(s): 鈴木英里子、宮田真路、岩尾康宏、佐藤ちひろ、北島健
  • Organizer: 第78回日本生化学中部支部例会・シンポジウム
  • Place of Presentation: 名古屋
  • Year and Date: 2014-05-24
• [Book] Handbook of Glycosyltransferases and related genes (2015)
  • Author(s): Sato, C.
  • Total Pages: 5
  • Publisher: Springer
• [Book] Glycoscience: Biology and Medicine (2015)
  • Author(s): Hane, M., Kitajima, K., and Sato, C.
  • Total Pages: 5
  • Publisher: Springer
• [Book] Glycoscience: Biology and Medicine (2015)
  • Author(s): 11.Sato, C., and Kitajima, K.
  • Total Pages: 7
  • Publisher: Springer
• [Remarks] Enzyme assay of polysialyltransferase
  • URL: http://jcggdb.jp/GlycoPOD/protocolShow.action?nodeId=t67
• [Remarks] 名古屋大学生物機能開発利用研究センター・動物細胞機能
  • URL: http://bbc.agr.nagoya-u.ac.jp/~acf/index.html