JP5684452B2 - 光干渉測定法により解剖学的構造に関連する情報を評価するためのシステム、方法及びソフトウエア装置 - Google Patents
光干渉測定法により解剖学的構造に関連する情報を評価するためのシステム、方法及びソフトウエア装置 Download PDFInfo
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Description
本出願は、2005年4月28日付で出願された米国特許出願第60/676,362号からの優先権の利益に基づき及びそれを主張する。その全ての開示は、引用文献により本明細書中に組み込まれる。
本発明は、国立衛生研究所により与えられた契約No.RO1 CA 103767の下でのアメリカ合衆国政府の支援によりなされた。従って、アメリカ合衆国政府は、本発明において特定の権利を有する。
本発明は、光干渉測定法(optical coherence ranging technique)により情報を評価する光干渉測定により解剖学的構造に関連する情報を評価するための、例えば生体から得られた顕微鏡画像を読み取るためのシステム、方法及びソフトウエア装置に関する。
胃食道逆流性疾患(GERD)は、発生率が増大し、食道特殊腸上皮化生(SIM)(一般的にバレット食道(BE)として知られている)の発達に関する周知の危険因子であって、R.J.Loffeld等の「Rising incidence of reflux oesophagitis in patients undergoing upper gastrointestinal endoscopy」(Digestion,2003,Vol.68(2−3)pp.141−4)に記載されている。C.Winters,Jr.等の「Barrett’s esophagus.A prevalent,occult complication of gastroesophageal reflux disease.Gastroenterology」(1987,Vol.92(1),pp.118−24)で論じされているように、慢性GERDを有する患者において、SIMの有病率は10〜15%もの高さであると推定された。J.Lagergren等の「Symptomatic gastroesophageal reflux as a risk factor for esophageal adenocarcinoma」(N Engl J Med,1999,Vol.340(11),pp.825−31)に記載されているように、GERDの再発及び重い症状を有する患者に関して、20年間にわたる腺癌の発達に対する調整オッズ比は、それぞれ7.7及び43.5である。更に、W.J.Blot等の「Rising incidence of adenocarcinoma of the esophagus and gastric cardia」(Jama,1991,Vol.265(10),pp.1287−9);P.Bytzer等の「Adenocarcinoma of the esophagus and Barrett’s esophagus:a population−based study」(Am J Gastroenterol,1999,Vol.94(1),pp.86−91);及び、S.S.Devesa等の「Changing patterns in the incidence of esophageal and gastric carcinoma in the United States」(Cancer,1998,Vol.83(10),pp.2049−53)で論じられているように、食道腺癌及び近位の胃(胃噴門)癌の発生率は、この30年間で急速に増大した。
BEが診断された場合、HGDを検出するための定期的な内視鏡サーベイランスが推奨され得る。P.Sharma等の「A critical review of the diagnosis and management of Barrett’s esophagus:the AGA Chicago Workshop」(Gastroenterology,2004,Vol.127(1),pp.310−30)に記載されているように、HGDを有する患者における腺癌の高い発生率(46ヶ月間で25%)を示す観測結果から、これらの推奨は進展し得る。D.S.Levine等の「An endoscopic biopsy protocol can differentiate high−grade dysplasia from early adenocarcinoma in Barrett’s esophagus」(Gastroenterology,1993,Vol.105(1),pp.40−50)で論じられているように、HGDのサーベイランスのための現在のガイドラインは、バレット部分の軸長に沿った2センチメーター毎の四象限生検を含み得る。しかし、G.S.Dulaiの「Surveying the case for surveillance」(Gastroenterology,2002,Vol.122(3),pp.820−823);G.W.Falk等の「Surveillance of patients with Barrett’s esophagus for dysplasia and cancer with balloon cytology」(Gastroenterology,1997,Vol.112(6),pp.1787−1797);及び、J.M.Streitz等の「Endoscopic surveillance of Barrett’s esophagus.Does it help?」(Journal of Thoracic and Cardiovascular Surgery,1993,Vol.105,pp.383−388)で論じられているように、サーベイランス内視鏡の正確性は、サンプリング誤差により限定され得る。J.W.van Sandick等の「Impact of endoscopic biopsy surveillance of Barrett’s oesophagus on pathological stage and clinical outcome of Barrett’s carcinoma」(Gut,1998,Vol.43(2),pp.216−22);J.M.Inadomi等の「Screening and surveillance for Barrett esophagus in high−risk groups:a cost−utility analysis」(Ann Intern Med,2003,VoI.138(3),pp.176−86);D.Provenzale等の「Barrett’s esophagus:a new look at surveillance based on emerging estimates of cancer risk」(Am J Gastroenterol,1999,Vol.94(8),pp.2043−53);及び、A Sonnenberg等の「Medical decision analysis of endoscopic surveillance of Barrett’s oesophagus to prevent oesophageal adenocarcinoma」(Aliment Pharmacol Ther,2002,Vol.16(1),pp.41−50)に記載されているように、BEに対する最適なサーベイランス及びスクリーニング・ストラテジーは議論されているが、多くの費用効果分析は、主な決定要因としての内視鏡の頻度及びコストに焦点を合わせている。
光干渉断層法(OCT)は、例えば近赤外光を用いて、胃腸粘膜の高分解能(10μmの軸方向分解能)の断面像を作成することのできる光学的画像診断法である。画像は、可視化された物質の特性に関連する光反射性に基づいて、構築することができる。S.Brand等の「Optical coherence tomography in the gastrointestinal tract」(Endoscopy,2000,Vol.32(10),pp.796−803)に記載されているように、OCT技術は、顕微鏡スケールの構造、例えば粘膜層、「窩(pit)及び腺」形態、及び腺状構造を同定するのに用いることができる。例えば、J.M.Poneros等の「Diagnosis of specialized intestinal metaplasia by optical coherence tomography」(Gastroenterology,2001,Vol.120(1),pp.7−12)で議論されているように、OCT法は、SIMを鱗状基底部及び腔粘膜から区別することができるが、胃噴門部をSIMとして誤って同定し得る。
異形成SIMの腺は、出芽、分岐、及び内腔陥入を有し、輪郭が込み合い、変形し、不規則であり得る。篩状腺、嚢状拡張、及び壊死片は、重度の異形成において同定される可能性が高い。
非異形成SIMは、最も大きな程度の表面成熟を有し、一方HGDは、最小の表面成熟を有し得る。高度の表面成熟は、表面の低い核対細胞質比率を暗示し、一方、低度の表面成熟は、高い表面の核対細胞質比率を示唆する。
異形成上皮を有する細胞は、一般的に、不規則な核膜、小嚢(異種)クロマチン、及び核極性の損失と共に、拡大した過色素性の核を含む。
炎症は、独立して変形した腺構造及び核異型を生じ得るので、炎症は、異形成の診断における交絡因子である。構造的及び核異型が炎症の結果であり得る場合は、異形成に関して不明確(IND)であると呼ばれる。組織学によるこの診断に対する観測者間同意は、アーチファクトが確定診断を提供するのに必要とされる特徴を不明瞭にする場合、又は疾患スペクトラムの異なる目的からの多くの基準が同時に存在する場合にしばしば留保されるので、低い(κ=0.14)(Montgomery等の「Reproducibility of the diagnosis of dysplasia in Barrett esophagus:a reaffirmation」(Human Pathology.2001,Vol.32,pp.368−378)に記載されている)。
本発明の1つの典型的な目的は、先行技術のシステム(例えば、上記に記載したもの)の特定の欠陥及び欠点を克服し、ケア組織病理の標準に相当する診断法を提供するための光学的生検画像を読み取るための典型的なシステム、方法及びソフトウエア装置を提供することである。本発明の別の典型的な目的は、光学的生検画像のための典型的なスコアリングシステム及び方法を提供することである。本発明の更に別の目的は、光学的生検画像から組織病理学的診断を得るための典型的なシステム、方法及びソフトウエア装置を提供することである。
図1は、本発明の光学的生検画像から診断を提供するための光学的生検スコアリングシステム/手順を生み出すための方法の典型的な実施態様を示す。図1に示されるこの典型的な方法は、段階100で一連の光学的生検画像を撮ること、及び段階110で切除生検染色組織病理スライド又はその画像を得ること、及び双方に対して共通する特徴を決定すること(段階120)を含む。特徴は、2つの画像セットの相互関係を比較することにより決定した形態学的特徴であることができる。特徴は、光学的生検画像及び組織病理の双方で同定することのできる個々の構造、パターン、強度、又は対応する病理組織構造に基づいた光学的生検画像構造の読み取りを含んで成ることができる。この特徴の例としては、上皮構造、上皮層、縁、腺の形状、不規則な腺の特徴、上皮成熟、核密度などが挙げられる。
i.典型的な設計
本発明の典型的な実施態様に関する典型的な試験は、盲式前向き試験であった。その主な目的は、SCJにおける腸上皮化生の分化に関するOCT画像特徴を同定することであった。ルーチンの外来上部消化管内視鏡検査を受ける患者は、試験に参加するように要求された。SCJのOCT画像は、内視鏡検査手順の間に得られた。2人の病理学者は、各生検標本を調査し、以下の組織タイプの存在に留意した:胃又は酸分泌噴門、鱗状粘膜、膵臓化生。腸上皮化生の存在は、杯細胞の存在により留意された。腸上皮化生の画像特徴は、既知の組織タイプの生検相関画像を含む、OCTアトラス「トレーニング・セット」を作り出し及び調査することにより決定した。その後、これらの特徴を、未知の組織タイプの「検証セット」に前向きに適用した。腸上皮化生の診断のための画像基準の感度、特異性、及び再現性を決定した。
本発明の典型的な実施態様に利用し、試験において用いることのできる典型的なOCT装置は、J.M.Poneros等の「Diagnosis of specialized intestinal metaplasia by optical coherence tomography」(Gastroenterology,2001,Vol.120(1),pp.7−12)、及びJ.M.Poneros等の「Optical coherence tomography of the biliary tree during ERCP」(Gastrointest Endosc,2002,Vol.55(1),pp.84−8)に記載されている。例えば、光源中心波長は1300nmで提供され、組織に投射する光強度は5.0mWであった。光源のスペクトルバンド幅は70nmであり、10μmの距離分解能を提供した。カテーテル径は、2.5mmであった。画像は、5.5mm(1000ピクセル)の長さ及び2.5mm(500ピクセル)の深さの縦方向の大きさを有する線形面において獲得された。画像獲得の間、フレームを1秒あたり2つの速度で記録し、参照のために連続的に番号を付ける。イメージングビームと同時の可視照準レーザーは、内視鏡医が、画像獲得を受ける粘膜の部位を特定するのを可能にし、画像化部位の生検相関を容易にした。
採用された被験者は、ルーチンの上部消化管内視鏡検査を受ける患者、及び胃食道接合部において既知の短い(<1cm)部分の腸上皮化生を有する患者を含む。3.8mmの器具チャネルを有する標準的な胃鏡(Pentax,Model EG 3470K、日本、東京)を用いた。
手順の前に、書面によるインフォームド・コンセントを得た。十分な鎮静及び口腔咽頭麻酔が達成された後に、上部消化管内視鏡検査を実施した。内視鏡医は、胃食道接合部又はバレット部分におけるSCJを同定した。OCTカテーテルプローブを、内視鏡の器具チャネルを通して導入し、SCJまで進めた。SCJのすぐ遠位で、OCT画像を獲得し、可視照準ビームによりマークした粘膜部位において記録し、そこでは、1つの大きな生検が得られた。画像化部位に対応するOCTフレームを記録した。患者毎に、2つの生検相関画像を得た。
生検標本を、10%のホルマリン中に置き、パラフィン中に埋め込み、ルーチンに処理し、ヘマトキシリン及びエオシンで染色した。
2人の病理学者は各生検標本を検査し、以下の上皮タイプの存在を決定した:胃噴門、鱗状粘膜、漿液膵臓化生、及び特殊腸上皮化生。この典型的な試験のために、鱗状粘膜及び酸分泌噴門(oxyntocardia)粘膜を、胃噴門として一緒に分類した。
20個の無作為に選んだSIMの生検相関画像及び20個の無作為に選んだ他の組織タイプの生検相関画像から成る画像アトラス「トレーニング・セット」を作成した。トレーニング・セットの画像のアトラスを検査し、SIMのための診断画像基準を決定した。その後、これらの基準を、残りのデータセットを含んで成る「検証セット」に前向きに適用した。検証セット中の全てのOCT画像を剥ぎ取って、情報を同定し無作為に混ぜた。
i.トレーニング・セット
扁平上皮を、腺を有しない層状上皮により区別した。図5Aは、水平層状構造を実証する扁平上皮の典型的なOCT画像を示す。図5Bは、胃噴門の典型的なOCT画像を示し、これは、通常の垂直な「窩及び腺」構造、高度に散乱した上皮表面、及び相対的に悪い画像透過性を示す。スケールバー、500μm。胃噴門(図5B中で示される)は、「窩及び腺」形態の存在、通常の表面構造、高度に反射する上皮表面の存在、又は悪い画像透過性により特徴づけられた。
検証セットを含んで成る156個の生検相関画像のうち、悪い画質により36個が除かれ、全部で120個の部位を前向き分析した。表1は、検証セットに関する組織病理を詳述する。
i 典型的な試験設計
実施した典型的な試験は、盲式前向き試験であった。採用した被験者は、ルーチンの内視鏡サーベイランス、又はIMC若しくはHGDに関する確認生検を受ける、BEを有する患者であった。バレット上皮のOCT画像は、内視鏡検査の間に得られた。食道の生検相関OCT画像は観察され、組織診断に対して盲式の読取者により評価された。各画像に関して、表面成熟及び腺構造に関するスコアをまとめ、「異形成指数」を確立した。各生検標本は独立に観察され、合意診断が提供された。
本発明の典型的な実施態様のために利用し、試験中で用いることのできる典型的なOCT装置は、J.M.Poneros等の「Diagnosis of specialized intestinal metaplasia by optical coherence tomography」(Gastroenterology,2001,Vol.120(1),pp.7−12)、及びJ.M.Poneros等の「Optical coherence tomography of the biliary tree during ERCP」(Gastrointest Endosc,2002,Vol.55(1),pp.84−8)に記載されている。例えば、光源中心波長は1300nmであり、組織に投射する光強度は5.0mWであった。光源のスペクトルバンド幅は70nmであり、10μmの距離分解能を提供した。カテーテル径は、2.5mmであった。画像は、5.5mm(1000ピクセル)の長さ及び2.5mm(500ピクセル)の深さの縦方向の大きさを有する線形面において獲得された。画像獲得の間、フレームを1秒あたり4つの速度で記録し、参照のために連続的に番号を付けた。イメージングビームと同時の可視照準レーザーは、内視鏡医が、画像獲得を受ける粘膜の部位を特定するのを可能にし、画像化部位の生検相関を容易にした。
被験者の手順の前に、インフォームド・コンセントを得た。サーベイランス内視鏡検査を受けるBEを有する患者、及び光線力学療法に関して評価されるHGC及びIMCの既知の診断を有する被験者を採用した。被験者は、ルーチンの意識下鎮静及び口腔咽頭麻酔を受けた。3.8mmの器具チャネルを有する標準的な内視鏡(Pentax,Model EG 3470K、日本、東京)を用いた。
十分な鎮静及び口腔咽頭麻酔の後に、上部消化管内視鏡検査を実施した。いったん内視鏡医は、胃食道接合部及びバレット部分を同定したら、OCTカテーテルプローブを、内視鏡の器具チャネルを通して導入し、バレット粘膜まで進めた。OCT画像を獲得し、集束ビームによりマークした粘膜部位において記録した。画像化部位に対応するOCTフレームを記録した。各画像化部位において、1つの大きな生検を実施した。
生検標本を、10%のホルマリン中に置き、パラフィン中に埋め込み、ルーチンに処理し、ヘマトキシリン及びエオシンで染色した。
vi 表面成熟の定義
OCTは、試料から返る光の強度を測定する。より高度に不均一な光学的屈折率を有する試料は、より強い光散乱、従ってより強いOCTシグナルを示す。ヒト組織の光学的特性を測定するために実施された以前の研究は、クロマチンの屈折率が細胞質[23]のものとは顕著に異なることを示した。このデータは、OCTシグナルが、核の大きさ及び密度の増大により増大するだろうことを示す。組織学的に、表面成熟は、表面における上皮の核対細胞質比の減少により部分的に特徴づけられる。従って、図9A〜Fに示すように、異形成の指標である不完全な表面成熟は、表面下シグナルと比較して高い表面OCTシグナルとして見られ得る。
OCT画像中の腺は、図9A〜9Fで示されるように、交互の低いOCTシグナル(細胞質)及び高いシグナル(核及び固有層)を有する線状構造として同定される。拡張した腺は、これらの図において、粘膜内の乏しい散乱空間として見られる。OCTによる腺の不規則性は、本明細書中で示されるように、これらの構造の不規則な大きさ、形状及び分布により特徴付けることができる。
例えば、OCT画像を剥ぎ取り、情報を同定し、無作為に混ぜて、画像のデータ・プールを作成する。この目的のために、IMCに一致する生検からの画像は、HGDの場合として含まれた。組織病理学的診断の調査をせずに、各OCT画像を調査し、以下のカテゴリーにおいて評価した:
A)表面成熟:0=表面下のOCTシグナルより弱い表面のOCTシグナル、1=表面下のOCTシグナルに等しい表面のOCTシグナル、2=表面下のOCTシグナルより強い表面のOCTシグナル
B)腺構造(0=不規則無し、正常な外見の腺構造;最小数の平らな拡張した腺;1=軽度の不規則性、腺は、より小さく密集し、又は大きく不規則な形状であった;拡張した腺は、より頻出し、密集していた;2=中程度/重度の不規則性、腺は分岐し、出芽していた;拡張した腺は、高度に非対称であり、又は腺の内腔中に残がいを含んでいた。
スピアマン相関係数(r)を計算して、各OCTで決定された組織病理学的特徴(表面成熟、腺構造、及び異形成指数)のスコアを、IMC/HGDの診断及び異形成(IMC/HGD、LGD、IGD)と比較した。IMC/HGD及び異形成(IMC/HGD、LGD、IGD)の診断のための異形成指数の感度及び特異性を計算した。統計は、SASソフトウエア(Statistical Analysis System,SAS Institute Inc.)バージョン8.0を用いた。p<0.05の値は、両側検定に関して統計的に有意であるとみなされた。
データセットは、58人の患者からの242個の生検相関画像から構成された。統計分析の前に、不十分な画質のため65個の画像を取り除いた。177個の残りの画像のうち、49個がIMC/HGD、15個がLGD、8個がIGD、100個がSIM、及び5個が胃粘膜の診断に対応した。65個の廃棄された画像のうち、20個がIMC/HGD、13個がLGD、2個がIGD、29個がSIM、及び1個が胃粘膜の診断に対応した。表3は、そのデータセットを含んで成る組織学的診断の分布を要約し、表面成熟、腺構造、及び異形成指数の平均OCTスコアを表示する。
表4は、各OCT画像特徴とIMC/HGDの診断との間のスピアマン相関係数を示す。それぞれの特徴とIMC/HGDの診断の間に正相関が存在した:[表面成熟(r=0.49、p<0.0001)、腺構造(r=.41、p<0.0001)、及び異形成指数(r=0.50、p<0.0001)]。3つの特徴のうち、異形成指数が最も高くIMC/HGDと相関した。
表4は、それぞれのOCTで決定された画像特徴と異形成の診断との間のスピアマン相関係数を示す。それぞれの特徴と異形成の診断との間に正相関が存在した[表面成熟(r=0.47、p<0.0001)、腺構造(r=0.44、p<0.0001)、及び異形成指数(r=0.50、p<0.0001)]。異形成と最も高い相関を有する画像特徴は、異形成指数であった。表6は、異形成指数のスコア>2が、異形成の診断に関して、72.0%(95%のCI、58%〜80%)の感度及び81.0%(95%のCI、72%〜88%)の特異性であることを実証する。
Claims (23)
- 解剖学的構造の少なくとも1つの部分に関連する第1の情報と解剖学的構造の少なくとも1つの部分に関連する第2の情報との間の関係を決定することにより、第3の情報を作成すること、ここで、上記第1の情報と第2の情報は、該情報間の関係を決定する前に予め得られたものであり、及び
所定の形態病理学的スコアリング基準及び第3の情報を用いて、少なくとも1つの画像を評価すること、
を含んで成る、解剖学的構造の少なくとも1つの部分に関連する少なくとも1つの画像を評価するための方法。 - 第1の情報又は第2の情報の少なくとも1つが、少なくとも1つの部分からの放射光と関連している、請求項1に記載の方法。
- 光が、少なくとも1つの部分から反射される、請求項2に記載の方法。
- 光が蛍光である、請求項2に記載の方法。
- 少なくとも1つの部分が、生体中で提供されたものである、請求項1に記載の方法。
- 解剖学的構造の少なくとも1つの部分が、顕微鏡用スライドの上に置かれる、請求項1に記載の方法。
- スライドが、ヘマトキシリン及びエオシン、マッソントリクロー、パパニコロー染色、ディフ・クイック、又は過ヨウ素酸シッフのうちの少なくとも1つにより染色される、請求項6に記載の方法。
- 第1の情報及び第2の情報が、解剖学的構造の少なくとも1つの部分のほぼ同一の場所に関して提供される、請求項1に記載の方法。
- 第3の情報が、第1の情報及び第2の情報に関連した物理的及び化学的構造に基づいて得られる、請求項1に記載の方法。
- 所定の形態病理学的スコアリング基準がハジット基準である、請求項9に記載の方法。
- 画像が、少なくとも1つの部分からの放射光と関連する、請求項1に記載の方法。
- 光が、少なくとも1つの部分から反射される、請求項11に記載の方法。
- 光が蛍光である、請求項11に記載の方法。
- 少なくとも1つの部分が、生体中で提供される、請求項11に記載の方法。
- 第1の情報又は第2の情報の少なくとも1つが、光干渉断層法システムにより得られる、請求項1に記載の方法。
- 第1の情報又は第2の情報の少なくとも1つが、スペクトルコード化共焦点顕微鏡システムにより得られる、請求項1に記載の方法。
- 解剖学的構造が、皮膚の下に存在する、請求項1に記載の方法。
- 第1の情報又は第2の情報の少なくとも1つが、共焦点顕微鏡システムにより得られる、請求項17に記載の方法。
- 第1の情報又は第2の情報の少なくとも1つが、反射共焦点顕微鏡システムにより得られる、請求項17に記載の方法。
- 第1の情報又は第2の情報の少なくとも1つが、光周波数ドメインイメージングシステムにより得られる、請求項1に記載の方法。
- スライドを抗体により染色する、請求項6に記載の方法。
- 所定の方法を実行する場合に、
a)解剖学的構造の少なくとも1つの部分に関連する第1の情報、及び 解剖学的構造の少なくとも1つの部分に関連する第2の情報を受け取り、
b)第1の情報と第2の情報との間の関係を決定することにより、第3の情報を作成し、及び
c)所定の形態病理学的スコアリング基準及び第3の情報を用いて、少なくとも1つの画像を評価する
ために設定された処理装置を含んで成る、解剖学的構造の少なくとも1つの部分に関連する少なくとも1つの画像を評価するための装置。 - 処理装置により実行される場合、解剖学的構造の少なくとも1つの部分に関連する第1の情報、及び 解剖学的構造の少なくとも1つの部分に関連する第2の情報を受け取るために処理装置を設定する第1の一連の指示、
処理装置により実行される場合、第1の情報と第2の情報との間の関係を決定することにより第3の情報を作成するために処理装置を設定する第2の一連の指示、及び
処理装置により実行される場合、所定の形態病理学的スコアリング基準及び第3の情報を用いて、少なくとも1つの画像を評価するために処理装置を設定する第3の一連の指示、
を含んで成る、解剖学的構造の少なくとも1つの部分に関連する少なくとも1つの画像を評価するためのソフトウエアシステム。
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EP1875436B1 (en) | 2009-12-09 |
JP2008541018A (ja) | 2008-11-20 |
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