參考用於說明的示例應用在下文中描述本發明的數個方面。應當理解的是，陳述許多具體細節、關係和方法來提供對本發明的充分理解。然而，在相關領域的普通技術人員將容易地認識到，可在不含一個或多個具體細節的情況下實施本發明或者可用其他方法來實施本發明。
本發明涉及新發現的生物標誌物(即RBM10)的突變和/或表現與西奧羅尼的療效、西奧羅尼的用藥和/或卵巢癌的治療效果之間的關係。本文所述生物標誌物提供用於評價西奧羅尼療效、指導西奧羅尼用藥和/或預測卵巢癌治療效果的方法。因此，本發明的一個實施方案代表生物標誌物的改進，所述腫瘤標誌物適用於評價西奧羅尼療效、指導西奧羅尼用藥和/或預測卵巢癌治療效果。在又一個實施方案中，本發明新發現的生物標誌物(即RBM10)可與本領域已知的一種或多種其它癌症標誌物(例如CEA、NSE、CA 19-9、CA 125、CA 72-4、PSA、proGRP、SCC、NNMT、VEGFR2、HER2、MISIIR、VEGFA、CD24)聯用，例如用於評價西奧羅尼療效、指導西奧羅尼用藥和/或預測卵巢癌治療效果或用於製備用於此目的的套組。
本發明通過以患者的無進展生存期(PFS)作為療效指標，驗證了RBM10基因變異與西奧羅尼療效的關聯性，對RBM10基因突變資訊的檢測，可以指導西奧羅尼的臨床用藥和評價其對腫瘤治療的療效，尤其適用於卵巢癌，特別適用於復發、晚期的卵巢癌。
在本發明中，“西奧羅尼的療效較佳、能夠使用西奧羅尼治療或者卵巢癌治療效果較好”尤其是指西奧羅尼能夠顯著延長患者例如卵巢癌患者的無進展生存期(PFS)。在某些實施方案中，本發明的套組用於評價西奧羅尼對治療卵巢癌的療效和/或指導西奧羅尼用於治療卵巢癌的用藥。
術語“樣品”意指已知或疑似表現或含有生物標誌物(即RBM10)或結合劑的材料，結合劑為例如對生物標誌物(即RBM10)有特異性的抗體。樣品可來源於生物來源(“生物樣品”)，例如組織(例如活組織檢查樣品)、提取物或包括細胞(例如腫瘤細胞)、細胞裂解物在內的細胞培養物和生物或生理流體，例如全血、血漿、血清、唾液、腦髓液、汗、尿液、乳汁、腹膜液等。獲自來源的樣品或在預處理以改進樣品特徵(例如從血液製備血漿、稀釋黏液等)後的樣品可直接使用。在本發明的某些方面，樣品是人生理流體，例如人血清。在本發明的某些方面，樣品是活組織檢查樣品例如經組織檢查獲得的腫瘤組織或細胞。在本發明的某些方面，樣品是惡性或正常組織樣品例如癌旁正常組織樣品。
可按照本發明進行分析的樣品包括臨床來源的多核苷酸。正如本領域技術人員應理解的是，靶多核苷酸可包括RNA，包括而不限於細胞總RNA、聚(A)+信使RNA(mRNA) 或其部分、胞質mRNA或由cDNA轉錄的RNA (即cRNA)。
可採用本領域已知方法，在一個或多個核苷酸上對靶多核苷酸進行可檢測標記。可檢測標記可以是而不限於發光標記、螢光標記、生物發光標記、化學發光標記、放射性標記和比色標記。
本文使用的術語“標誌物”指要用作分析患者實驗樣品的靶標的分子。這樣的分子靶標的實例是基因、蛋白或多肽。在本發明中用作標誌物的基因、蛋白或多肽預期包括所述基因或蛋白的天然存在的變體以及所述基因或蛋白或所述變體的片段，特別是免疫學上可檢測的片段。免疫學上可檢測的片段優選地包含所述標誌物多肽的至少6、7、8、10、12、15或20個連續胺基酸。本領域的技術人員可認識到，由細胞釋放的蛋白或存在於胞外基質中的蛋白可能受到損害(例如在炎症過程中)，且可被降解或切割成這樣的片段。某些標誌物以無活性形式合成，其可以隨後通過蛋白酶解來啟動。如熟練的技術人員將明白的，蛋白或其片段也可以作為複合物的部分而存在。這樣的複合物也可以用作本發明意義上的標誌物。標誌物多肽的變體由相同的基因編碼，但可能在其等電點(=PI)或分子量(=MW)或以上二者上有差異，例如作為可選的mRNA或mRNA前體加工的結果。變體的胺基酸序列與對應的標誌物序列具有95%、96%、97%、98%、99%或更高的同一性。另外，或在替代方案中，標誌物多肽或其變體可以攜帶轉譯後修飾。轉譯後修飾的非限制性實例是糖基化、醯化和/或磷酸化。
標誌物的表現也可通過檢測標誌物的轉譯(即，樣品中標誌物蛋白的檢測)來鑒定。適合於檢測標誌物蛋白的方法包括用於檢測和/或測量得自細胞或細胞提取物的蛋白的任何合適方法。這樣的方法包括，但不限於免疫印跡(如蛋白質印跡)、酶聯免疫吸附測定(ELISA)、放射免疫測定(RIA)、免疫沉澱、免疫組織化學和免疫螢光。用於檢測蛋白的特別優選的方法包括任何基於細胞的測定，包括免疫組織化學和免疫螢光測定。這樣的方法是本領域熟知的。
術語“受試者”、“患者”和“個體”在本文可互換使用，是指溫血動物，例如哺乳動物。該術語包括但不限於家畜、齧齒動物(例如大鼠和小鼠)、靈長類動物和人。優選該術語是指人。
RBM10(NG_012548.1)屬於含RNA基序結合蛋白家族，位於染色體Xp11.23，編碼一個含有930個胺基酸的核蛋白，並含有兩個RNA識別結構域(RRM)、兩個鋅指結構域和一個G-patch結構域。該基因也稱為DXS8237E、GPATC9、GPATCH9、S1-1、TARPS和ZRANB5。
術語“野生型”應根據本領域技術人員的一般理解來理解，並且表示呈天然存在的形式而沒有進行任何人工突變、核苷酸變換或胺基酸修改的核酸或胺基酸序列。
術語“突變型”應根據本領域技術人員的一般理解來理解。如果核酸序列與其自然或天然核酸序列相比，在其核酸序列中含有至少一個核苷酸的添加、缺失或取代，即如果它含有核酸突變，則將其稱為“突變的”。如果胺基酸序列與其自然或天然胺基酸序列相比，在其胺基酸序列中含有至少一個增加的、缺失的或取代的胺基酸，即如果它含有胺基酸突變，則將其稱為“突變的”。
在本發明中，RBM10基因或其mRNA的變異是指野生型RBM10基因或其mRNA中的突變，其包括一個或多個核苷酸的添加、缺失和取代以及拷貝數的變化(包括擴增和缺失)。所述核苷酸可以是編碼區或非編碼區的核苷酸。所述編碼區可以是編碼RNA識別結構域(RRM)、鋅指結構域和/或G-patch結構域的核苷酸序列。所述RBM10基因或其mRNA的變異可以導致RBM10基因表現水準的上升或下降和/或拷貝數的擴增或缺失。例如水準或拷貝數分別為對照或標準的至少約1.1、1.25、1.5、2、3、4、5、6、7、8、9或10倍或更多倍或者為對照或標準的至多約1/1.1、1/1.25、1/1.5、1/2、1/3、1/4、1/5、1/6、1/7、1/8、1/9或1/10或更少。拷貝數擴增或缺失可通過本領域周知的技術檢測，例如全基因測序。
在本發明中，RBM10基因編碼的蛋白或蛋白片段的變異包括：1)野生型RBM10基因所編碼的蛋白或蛋白片段中的突變，其包括一個或多個胺基酸的增加、缺失或取代；以及2)野生型RBM10基因所編碼的蛋白的表現水準的變化。所述胺基酸可以是RNA識別結構域(RRM)、鋅指結構域和/或G-patch結構域的胺基酸。所述蛋白片段是指與全長天然蛋白相比具有胺基末端缺失、羧基末端缺失和/或中間缺失的多肽。所述片段還可含有與天然蛋白相比經修飾的胺基酸。在某些實施方案中，片段長度為約5-500個胺基酸。例如，片段長度可為至少5、6、8、10、14、20、50、70、100、110、150、200、250、300、350、400或450個胺基酸。在一個實施方案中，所述片段是免疫學上可檢測的片段，其優選地包含標誌物多肽的至少6、7、8、10、12、15或20個連續胺基酸。所述蛋白表現水準的變化是指與對照或標準的表現水準相比，至少為對照或標準的表現水準的至少約1.1、1.25、1.5、2、3、4、5、6、7、8、9或10倍或更多倍或者為對照或標準的表現水準的至多約1/1.1、1/1.25、1/1.5、1/2、1/3、1/4、1/5、1/6、1/7、1/8、1/9或1/10或更少。
術語“多肽”和“蛋白質”在本文可互換使用，表示通過共價和/或非共價鍵連接的胺基酸的至少一個分子鏈。該術語包括肽、寡肽和蛋白質及多肽的轉譯後修飾，例如糖基化、乙醯化、磷酸化等。蛋白質片段、類似物、突變蛋白質或變體蛋白質、融合蛋白等也包括在該術語的含義中。
在某些實施方案中，如本文所用的測定“蛋白表現水準”、“基因表現”或“基因表現水準”包括但不限於測定相應的RNA、蛋白或者肽水準(或其組合)。本發明不限於測定蛋白、肽或RNA水準的具體方法和試劑，所有這些方法和試劑是本領域熟知的。
用於測定樣品中蛋白質的量或濃度的方法為技術人員所知。所述方法包括放射性免疫測定、競爭性結合測定、蛋白質印跡分析和ELISA測定。對於使用抗體的方法，單株和多株抗體都適用。所述抗體對於蛋白質、蛋白表位或蛋白片段可為免疫學上特異的。
術語“寡核苷酸”是指任何長度的核苷酸的多聚體形式，為核糖核苷酸或去氧核糖核苷酸。該術語包括雙鏈和單鏈DNA和RNA，例如多核苷酸的甲基化或帽化等修飾形式和未修飾形式。術語“多核苷酸”和“寡核苷酸”在本文可互換使用。寡核苷酸可但非必需包括其它編碼或非編碼序列，或者它可以但不一定與其它分子和/或載體或支援材料連接。用於本發明方法或套組的寡核苷酸可具有適於具體方法的任何長度。在某些應用中，該術語是指反義核酸分子(例如處於與編碼本發明癌症標誌物(例如RBM10)的有義多核苷酸相反方向的mRNA或DNA鏈)。
用於本發明的寡核苷酸包括互補核酸序列和與這些序列基本相同的核酸，並且還包括因遺傳密碼簡並而不同於核酸序列的序列。可用於本發明的寡核苷酸還包括在嚴格條件下、優選高嚴格性條件下與寡核苷酸癌症標誌物核酸序列雜交的核酸。
核苷酸雜交測定是本領域熟知的。雜交測定程式和條件將根據應用而變化並依據已知的通用結合方法選擇，參見例如J. 薩姆布魯克等，分子克隆：實驗指南(第三版. 科學出版社，2002)；以及Young和Davis，P.N.A.S，80: 1194 (1983)。進行重複和受控雜交反應的方法和設備已經描述於美國專利號5,871,928、5,874,219、6,045,996、6,386,749和6,391,623中，其各自通過引用結合到本文中。
在某些情況下，可能需要擴增樣品。基因組樣品可通過各種機制擴增，其中一些機制可採用PCR。樣品可在陣列上擴增。參見，例如美國專利號6,300,070和美國專利申請系列號09/513,300。
其它合適的擴增方法包括連接酶鏈反應(LCR) (如Wu和Wallace，Genomics 4，560 (1989)、Landegren等Science 241、1077 (1988)和Barringer等Gene 89:117 (1990))、轉錄擴增(Kwoh等，Proc. Natl. Acad. Sci. USA 86，1173 (1989)和WO88/10315)、自維持序列複製(Guatelli等，Proc. Nat. Acad. Sci. USA，87，1874 (1990)和WO90/06995)、目標多核苷酸序列的選擇性擴增(美國專利號6,410,276)、共有序列引發的聚合酶鏈反應(CP-PCR) (美國專利號4,437,975)、任意引發的聚合酶鏈反應(AP-PCR)(美國專利號5,413,909、5,861,245)和基於核酸的序列擴增(NABSA) (參見美國專利號5,409,818、5,554,517和6,063,603，其各自通過引用結合到本文中)。
可用於檢測RBM10表現水準和/或拷貝數的試劑是本領域眾所周知的。適用於本發明的這種試劑可市購獲得或通過本領域技術人員熟知的方法常規地製得。
術語“結合劑”是指例如與本發明生物標誌物(RBM10)特異性結合的多肽、抗體、核糖體或適體等物質。如果物質以可檢測的水準與本發明生物標誌物起反應，而不與含有無關序列或不同多肽的序列的肽可檢測地起反應，則它與本發明生物標誌物“特異性結合”。可採用本領域技術人員可容易進行的ELISA來評價結合性質。
結合劑可以是含或不含肽組分的核糖體、RNA或DNA分子或多肽。結合劑可以是包含多肽生物標誌物序列、其肽變體或這類序列的非肽模擬物的多肽。
適體包括與核酸和蛋白質結合的DNA或RNA分子。與本發明標誌物結合的適體可在無需過多實驗的情況下利用常規技術產生。[例如參見下列描述適體體外選擇的出版物：Klug等, Mol. Biol. Reports 20:97-107(1994)；Wallis等, Chem. Biol. 2:543-552(1995)；Ellington, Curr. Biol. 4:427-429(1994)；Lato等, Chem. Biol. 2:291-303(1995)；Conrad等, Mol. Div. 1:69-78(1995)；以及Uphoff等, Curr. Opin. Struct. Biol. 6:281-287(1996)]。
用於本發明的抗體包括但不限於合成抗體、單株抗體、多株抗體、重組抗體、抗體片段(例如Fab、Fab'、F(ab')2)、dAb(結構域抗體；參見Ward等，1989，Nature，341:544-546)、抗體重鏈、胞內抗體、人源化抗體、人抗體、抗體輕鏈、單鏈Fv(scFv)(例如包括單特異性、雙特異性等)、抗獨特型(ant-Id)抗體、包含抗體部分的蛋白質、嵌合抗體(例如含有鼠抗體的結合特異性但其中其餘部分是人來源的抗體)、衍生物例如酶綴合物或標記的衍生物、雙鏈抗體、線性抗體、二硫鍵連接的Fv(sdFv)、多特異性抗體(例如雙特異性抗體)、上述任一種的表位結合片段和包含所需特異性的抗原識別部位的免疫球蛋白分子的任何其它修飾構型。抗體包括任何類型(例如IgA、IgD、IgE、IgG、IgM和IgY)、任何類別(例如IgG1、IgG2、IgG3、IgG4、IgA1和IgA2)或任何亞類(例如IgG2a和IgG2b)的抗體，抗體不必是任何特定的類型、類別或亞類。在本發明的某些實施方案中，抗體是IgG抗體或其類別或亞類。抗體可來自任何動物來源，包括鳥類和哺乳動物(例如人、鼠、驢、綿羊、兔、山羊、豚鼠、駱駝、馬或雞)。
例如，用於本發明的抗體可市購自例如Invitrogen(貨號PA5-83253、PA5-40331、PA5-40330等)、Abcam (例如貨號為ab224149的抗體)等。或者，所述抗體可通過本領域周知的重組方法製備。在一些實施方案中，所述抗體是單株抗體。對於單株抗體的製備參見例如Kohler等(1975) Nature 256:495-497；Kozbor等(1985) J. Immunol Methods 81:31-42；Cote等(1983) Proc Natl Acad Sci 80:2026-2030和Cole等(1984) Mol Cell Biol 62:109-120。
本發明的套組可以通過本領域常規方法製備。套組可包含實施本發明方法所用的材料或試劑(包括用於檢測RBM10基因或其mRNA或其編碼的蛋白或蛋白片段的試劑)。套組可以包括儲存反應試劑(例如在合適容器中的引物、dNTP、酶等)和/或支持材料(例如緩衝液、實施檢測的說明書等)。例如，套組可以包括一個或多個含有相應反應試劑和/或支持材料的容器(例如盒子)。這樣的內容物可一起或分開遞送給既定的接受者。作為一個實例，套組可含有用於檢測RBM10基因或其mRNA或其編碼的蛋白或蛋白片段的試劑、緩衝液以及使用說明書。套組還可含有聚合酶和dTNP等。套組還可含有用於質控的內標、陽性和陰性對照等。套組還可包含用於從樣品製備核酸例如DNA的試劑。以上實例不能理解為限制適用於本發明的套組及其內容物。
微陣列是指具有平坦表面的固相支援體，其具有核酸陣列，陣列中的各個成員包含固定在空間上確定的區域或位元點上的寡核苷酸或多核苷酸的相同的拷貝，所述區域或位元點不與陣列中的其它成員的區域或位元點重疊；也就是說，所述區域或位元點在空間上是離散的。此外，空間上確定的雜交位點可為“可定址的”，因為其位置及其固定化的寡核苷酸的身份是已知或預先確定的(例如在其使用前是已知或預先確定的)。通常寡核苷酸或多核苷酸為單鏈，並通常由5'-端或3'-端與固相支援體共價連接。微陣列中含有非重疊區的核酸的密度通常大於100/cm2
，更優選大於1000/cm2
。微陣列技術公開於例如以下參考文獻中：Schena編輯的Microarrays: A Practical Approach (IRL Press, Oxford, 2000)；Southern, Current Opin. Chem. Biol., 2:404-410，1998，其全部內容通過引用結合到本文中。
本發明公開了RBM10基因的用途，本領域技術人員可以借鑒本文內容，適當改進工藝參數實現。特別需要指出的是，所有類似的替換和改動對本領域技術人員來說是顯而易見的，它們都被視為包括在本發明。本發明所述用途已經通過較佳實施例進行了描述，相關人員明顯能在不脫離本發明內容、精神和範圍內對本文所述用途進行改動或適當變更與組合，來實現和應用本發明技術。
以下就本發明所提供的RBM10基因的用途做進一步說明。實施例 1 ：
西奧羅尼單藥治療復發難治的卵巢癌Ib期臨床試驗試驗藥物：
西奧羅尼膠囊，規格：5mg、25mg。由深圳微芯生物科技股份有限公司生產。給藥方案：
西奧羅尼膠囊按50mg/天、QD給藥(對體重或體表面積不做調整)。每天上午空腹服用，用水送服，完整吞服整粒膠囊。連續給藥28天為1個治療週期，每個治療周期間無間隔。病例數：
入組25例，其中有療效評價的患者23例。入選標準：
1.年齡≥18歲，≤70歲，女性；
2.經組織學確診的上皮性卵巢癌、輸卵管癌或原發性腹膜癌；
3.受試者需接受過含鉑類化療方案，即
a) 鉑類耐藥者(含鉑類化療方案結束後≤6個月疾病進展或復發)，應接受過至少2個不同化療方案後疾病進展或復發；
b) 若鉑類敏感者(含鉑類化療方案結束後＞6個月疾病進展或復發)，應接受過至少2個化療方案後疾病進展或復發，或者受試者拒絕接受再次化療；
4.根據RECIST1.1標準，至少有一個可測量的靶病灶；
5.ECOG體力評分0-1分；
6.距離前次的化療、放療、靶向治療、免疫治療或研究藥物治療結束需間隔4周以上，如化療方案包含絲裂黴素，間隔應在6周以上；
7.主要器官功能符合以下標準：
血常規：中性粒細胞絕對值≥1.5×109
/L，血小板≥90×109
/L，血紅蛋白≥90g/L；
血生化：總膽紅素≤1.5倍正常值參考範圍上限(ULN)，AST、ALT≤2倍ULN(肝轉移病例：≤5倍ULN)；血清肌酐≤1.5倍ULN；
凝血功能：凝血酶原時間-國際標準化比率(PT-INR)≤1.5倍ULN。
8.預期生存時間≥3個月；
9.自願簽署書面知情同意書。治療計畫：
試驗受試者每天口服西奧羅尼膠囊50mg一次，每28天為一個治療週期，治療周期間無停藥間隔期。整個試驗期間，所有受試者均持續治療直至出現以下任一情況(以先發生者為準)：疾病進展、不能耐受的毒性反應、死亡、撤回知情同意或失訪。療效評估：
根據RECIST1.1標準，分別在基線期以及治療後第4週末評估一次，後續每8周重複進行，直到疾病進展。腫瘤影像學檢查包括頸部、胸部、全腹、盆腔CT或MRI，其他部位檢查根據臨床指征，有需要時進行，病灶基線和後續評估測量應採用同樣的技術和方法。安全性評估：
包括體格檢查、生命體征、ECOG體能評分、血常規、尿常規、12導聯ECG、血生化、電解質、凝血功能、心肌酶、肌鈣蛋白、TSH、FT3、FT4、澱粉酶、超聲心動圖、24小時尿蛋白定量(必要時)、不良事件。生物標誌物研究：
在受試者首次服用西奧羅尼前和疾病進展時取外周血10毫升，對血漿游離腫瘤DNA(ctDNA)和白細胞提取DNA(對照)進行基因序列的檢測，共包括548個腫瘤相關基因，檢測結果包括基因突變和拷貝數異常。臨床試驗結果：
25例受試者中，有2例無基線後療效評估結果，因此可評價病例為23例。23例可評價受試者中，試驗期間最佳療效為：2例(8.7%)PR(其中1例在後續評估中得到確認)，14例(60.9%)SD，7例(30.4%)PD，確認的ORR為4.3%，未確認ORR為8.7%。23例可評價受試者中，有3例鉑敏感，20例鉑耐藥。2例PR受試者均為鉑耐藥受試者，確認的緩解為1例(5.0%)。結果顯示西奧羅尼單藥治療卵巢癌有效。
對已評價患者血漿游離腫瘤DNA(ctDNA)檢測分析，對548個腫瘤相關基因進行了療效相關生物標誌物的伴隨研究。檢測結果選取了所有突變發生率0.4%以上的基因，以患者的無進展生存期(PFS)作為療效指標，分析腫瘤相關基因異常與西奧羅尼療效的關聯性。結果顯示，在548個腫瘤相關基因中，發現了一個與西奧羅尼療效有顯著相關性的基因突變，為RBM10基因。可評價23例受試者中RBM10突變7例(30.4%，這些突變包括核苷酸的添加、缺失和取代以及拷貝數的變化)的中位PFS為232天，RBM10野生型16例(69.6%)的中位PFS為106天，兩組具有顯著差異，提示RBM10基因與西奧羅尼治療卵巢癌療效相關，可以成為評價療效的潛在生物標誌物。結果參見表1和圖1。
以上所述僅是本發明的優選實施方式，應當指出，對於本技術領域的普通技術人員來說，在不脫離本發明原理的前提下，還可以做出若干改進和潤飾，這些改進和潤飾也應視為本發明的保護範圍。Several aspects of the invention are described below with reference to example applications for illustration. It should be understood that numerous specific details, relationships and methods are set forth to provide a thorough understanding of the present invention. However, one of ordinary skill in the relevant art will readily recognize that the invention may be practiced without one or more of the specific details or in other ways. The present invention relates to the relationship between the mutation and/or expression of a newly discovered biomarker (ie RBM10) and the efficacy of cioronib, the administration of cioronib and/or the therapeutic effect of ovarian cancer. The biomarkers described herein provide methods for evaluating the efficacy of cioronib, guiding the administration of cioronib, and/or predicting the effect of treatment in ovarian cancer. Thus, one embodiment of the present invention represents an improvement in biomarkers suitable for evaluating the efficacy of cioronib, guiding the administration of cioronib and/or predicting the effect of ovarian cancer treatment. In yet another embodiment, the newly discovered biomarkers of the present invention (ie, RBM10) may be combined with one or more other cancer markers known in the art (eg, CEA, NSE, CA 19-9, CA 125, CA 72- 4. Combination of PSA, proGRP, SCC, NNMT, VEGFR2, HER2, MISIIR, VEGFA, CD24), for example, for evaluating the efficacy of cioronib, guiding the administration of cioronib and/or predicting the treatment effect of ovarian cancer or for A kit is prepared for this purpose. The invention verifies the correlation between the RBM10 gene variation and the curative effect of cioroni by taking the patient's progression-free survival (PFS) as the curative effect index, and the detection of the RBM10 gene mutation information can guide the clinical medication and treatment of cioroni. To evaluate its curative effect on tumor treatment, especially for ovarian cancer, especially for recurrent and advanced ovarian cancer. In the present invention, “the curative effect of cioronib is better, the treatment effect of cioronib can be better, or the treatment effect of ovarian cancer is better” especially refers to that cioronib can significantly prolong the progression-free survival period of patients such as ovarian cancer patients (PFS). In certain embodiments, the panels of the present invention are used to evaluate the efficacy of cioronib for the treatment of ovarian cancer and/or to guide the administration of cioronib for the treatment of ovarian cancer. The term "sample" means a material known or suspected to express or contain a biomarker (ie RBM10) or a binding agent, eg an antibody specific for the biomarker (ie RBM10). Samples may be derived from biological sources ("biological samples"), such as tissues (eg, biopsy samples), extracts or cell cultures including cells (eg, tumor cells), cell lysates, and biological or physiological fluids, such as Whole blood, plasma, serum, saliva, cerebrospinal fluid, sweat, urine, milk, peritoneal fluid, etc. Samples obtained from sources or after pretreatment to improve sample characteristics (eg, preparation of plasma from blood, dilution of mucus, etc.) can be used directly. In certain aspects of the invention, the sample is a human physiological fluid, such as human serum. In certain aspects of the invention, the sample is a biopsy sample such as tumor tissue or cells obtained by tissue examination. In certain aspects of the invention, the sample is a malignant or normal tissue sample such as a paracancerous normal tissue sample. Samples that can be analyzed in accordance with the present invention include polynucleotides of clinical origin. As will be understood by those of skill in the art, target polynucleotides can include RNA, including, but not limited to, total cellular RNA, poly(A)+ messenger RNA (mRNA) or a portion thereof, cytoplasmic mRNA, or RNA transcribed from cDNA ( i.e. cRNA). The target polynucleotide can be detectably labeled on one or more nucleotides using methods known in the art. Detectable labels can be, without limitation, luminescent, fluorescent, bioluminescent, chemiluminescent, radioactive, and colorimetric labels. The term "marker" as used herein refers to a molecule to be used as a target for analyzing a patient's experimental sample. Examples of such molecular targets are genes, proteins or polypeptides. The genes, proteins or polypeptides used as markers in the present invention are intended to include naturally occurring variants of said genes or proteins as well as fragments of said genes or proteins or said variants, particularly immunologically detectable fragments . Immunologically detectable fragments preferably comprise at least 6, 7, 8, 10, 12, 15 or 20 contiguous amino acids of the marker polypeptide. One skilled in the art will recognize that proteins released by cells or present in the extracellular matrix may be damaged (eg, during inflammation) and may be degraded or cleaved into such fragments. Certain markers are synthesized in an inactive form, which can then be initiated by proteolysis. As the skilled artisan will appreciate, proteins or fragments thereof can also be present as part of a complex. Such complexes can also be used as markers in the sense of the present invention. Variants of marker polypeptides are encoded by the same gene, but may differ in their isoelectric point (=PI) or molecular weight (=MW) or both, e.g. as a result of alternative mRNA or pre-mRNA processing . The amino acid sequence of the variant is 95%, 96%, 97%, 98%, 99% or more identical to the corresponding marker sequence. Additionally, or in the alternative, marker polypeptides or variants thereof may carry post-translational modifications. Non-limiting examples of post-translational modifications are glycosylation, acylation and/or phosphorylation. The performance of the marker can also be identified by detecting translation of the marker (ie, detection of the marker protein in the sample). Methods suitable for detecting marker proteins include any suitable method for detecting and/or measuring proteins from cells or cell extracts. Such methods include, but are not limited to, immunoblotting (eg, Western blotting), enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), immunoprecipitation, immunohistochemistry, and immunofluorescence. Particularly preferred methods for detecting proteins include any cell-based assay, including immunohistochemistry and immunofluorescence assays. Such methods are well known in the art. The terms "subject", "patient" and "individual" are used interchangeably herein to refer to a warm-blooded animal, such as a mammal. The term includes, but is not limited to, livestock, rodents (eg, rats and mice), primates, and humans. Preferably the term refers to humans. RBM10 (NG_012548.1) belongs to a family of RNA motif-binding proteins, located on chromosome Xp11.23, encoding a 930 amino acid nucleoprotein with two RNA recognition domains (RRMs) and two zinc finger structures domain and a G-patch domain. This gene is also known as DXS8237E, GPATC9, GPATCH9, S1-1, TARPS and ZRANB5. The term "wild-type" is to be understood according to the general understanding of those skilled in the art and refers to a nucleic acid or amino acid sequence in its naturally occurring form without any artificial mutation, nucleotide change or amino acid modification. The term "mutant" should be understood according to the general understanding of those skilled in the art. A nucleic acid sequence is referred to as "mutated" if it contains at least one nucleotide addition, deletion or substitution in its nucleic acid sequence compared to its native or native nucleic acid sequence, ie if it contains a nucleic acid mutation. If the amino acid sequence contains at least one added, deleted or substituted amino acid in its amino acid sequence compared to its natural or native amino acid sequence, i.e. if it contains an amino acid mutation, then called "mutated". In the present invention, the variation of the RBM10 gene or its mRNA refers to the mutation in the wild-type RBM10 gene or its mRNA, which includes addition, deletion and substitution of one or more nucleotides and changes in copy number (including amplification and missing). The nucleotides may be nucleotides of a coding region or a non-coding region. The coding region may be a nucleotide sequence encoding an RNA recognition domain (RRM), a zinc finger domain and/or a G-patch domain. The variation of the RBM10 gene or its mRNA can lead to an increase or decrease in the expression level of the RBM10 gene and/or an amplification or deletion of the copy number. For example, the level or copy number is at least about 1.1, 1.25, 1.5, 2, 3, 4, 5, 6, 7, 8, 9 or 10 times or more of the control or standard, respectively or at most about 1 of the control or standard /1.1, 1/1.25, 1/1.5, 1/2, 1/3, 1/4, 1/5, 1/6, 1/7, 1/8, 1/9 or 1/10 or less. Copy number amplifications or deletions can be detected by techniques well known in the art, such as whole gene sequencing. In the present invention, the variation of the protein or protein fragment encoded by the RBM10 gene includes: 1) mutations in the protein or protein fragment encoded by the wild-type RBM10 gene, which include addition, deletion or substitution of one or more amino acids; and 2) changes in the expression level of the protein encoded by the wild-type RBM10 gene. The amino acid may be an amino acid of an RNA recognition domain (RRM), a zinc finger domain and/or a G-patch domain. The protein fragments refer to polypeptides having amino-terminal deletions, carboxy-terminal deletions and/or intermediate deletions compared to the full-length native protein. The fragments may also contain modified amino acids compared to the native protein. In certain embodiments, the fragments are about 5-500 amino acids in length. For example, fragments can be at least 5, 6, 8, 10, 14, 20, 50, 70, 100, 110, 150, 200, 250, 300, 350, 400, or 450 amino acids in length. In one embodiment, the fragment is an immunologically detectable fragment preferably comprising at least 6, 7, 8, 10, 12, 15 or 20 contiguous amino acids of the marker polypeptide. The change in the expression level of the protein refers to at least about 1.1, 1.25, 1.5, 2, 3, 4, 5, 6, 7, 8, 9 or 10 times or more or at most about 1/1.1, 1/1.25, 1/1.5, 1/2, 1/3, 1/4, 1/5, 1/6 of the performance level of a control or standard , 1/7, 1/8, 1/9 or 1/10 or less. The terms "polypeptide" and "protein" are used interchangeably herein to refer to at least one molecular chain of amino acids linked by covalent and/or non-covalent bonds. The term includes post-translational modifications of peptides, oligopeptides, and proteins and polypeptides, such as glycosylation, acetylation, phosphorylation, and the like. Protein fragments, analogs, muteins or variant proteins, fusion proteins, etc. are also included within the meaning of the term. In certain embodiments, determining "protein expression levels,""geneexpression," or "gene expression levels" as used herein includes, but is not limited to, determining corresponding RNA, protein, or peptide levels (or combinations thereof). The present invention is not limited to specific methods and reagents for measuring protein, peptide or RNA levels, all of which are well known in the art. Methods for determining the amount or concentration of protein in a sample are known to the skilled artisan. Such methods include radioimmunoassays, competitive binding assays, Western blot analysis and ELISA assays. For methods using antibodies, both monoclonal and polyclonal antibodies are suitable. The antibody may be immunologically specific for a protein, protein epitope or protein fragment. The term "oligonucleotide" refers to a polymeric form of nucleotides of any length, either ribonucleotides or deoxyribonucleotides. The term includes double- and single-stranded DNA and RNA, modified and unmodified forms such as methylation or capping of polynucleotides. The terms "polynucleotide" and "oligonucleotide" are used interchangeably herein. An oligonucleotide may, but need not, include other coding or non-coding sequences, or it may, but need not, be linked to other molecules and/or vectors or support materials. The oligonucleotides used in the methods or kits of the present invention can be of any length suitable for the particular method. In certain applications, the term refers to an antisense nucleic acid molecule (eg, an mRNA or DNA strand in the opposite orientation to a sense polynucleotide encoding a cancer marker of the invention (eg, RBM10)). Oligonucleotides for use in the present invention include complementary nucleic acid sequences and nucleic acids substantially identical to those sequences, and also include sequences that differ from nucleic acid sequences due to the degeneracy of the genetic code. Oligonucleotides useful in the present invention also include nucleic acids that hybridize to oligonucleotide cancer marker nucleic acid sequences under stringent conditions, preferably high stringency conditions. Nucleotide hybridization assays are well known in the art. Hybridization assay protocols and conditions will vary depending on the application and are chosen according to known general binding methods, see eg, J. Sambrook et al., Molecular Cloning: A Laboratory Guide (Third Edition. Science Press, 2002); and Young and Davis , PNAS, 80: 1194 (1983). Methods and apparatus for performing repetitive and controlled hybridization reactions have been described in US Pat. Nos. 5,871,928, 5,874,219, 6,045,996, 6,386,749, and 6,391,623, each of which is incorporated herein by reference. In some cases, it may be necessary to amplify the sample. Genomic samples can be amplified by various mechanisms, some of which can employ PCR. Samples can be amplified on the array. See, eg, US Patent No. 6,300,070 and US Patent Application Serial No. 09/513,300. Other suitable amplification methods include ligase chain reaction (LCR) (eg Wu and Wallace, Genomics 4, 560 (1989), Landegren et al. Science 241, 1077 (1988) and Barringer et al. Gene 89:117 (1990)), transcription Amplification (Kwoh et al., Proc. Natl. Acad. Sci. USA 86, 1173 (1989) and WO 88/10315), self-sustaining sequence replication (Guatelli et al., Proc. Nat. Acad. Sci. USA, 87, 1874 (1990) ) and WO90/06995), selective amplification of target polynucleotide sequences (US Pat. No. 6,410,276), consensus-primed polymerase chain reaction (CP-PCR) (US Pat. No. 4,437,975), arbitrarily primed polymerase chain reaction reaction (AP-PCR) (US Pat. Nos. 5,413,909, 5,861,245) and Nucleic Acid-Based Sequence Amplification (NABSA) (see US Pat. Nos. 5,409,818, 5,554,517 and 6,063,603, each of which is incorporated herein by reference). Reagents that can be used to detect RBM10 expression levels and/or copy number are well known in the art. Such reagents suitable for use in the present invention are commercially available or routinely prepared by methods well known to those skilled in the art. The term "binding agent" refers to substances such as polypeptides, antibodies, ribosomes or aptamers that specifically bind to the biomarkers of the invention (RBM10). A substance "specifically binds" to a biomarker of the invention if it reacts at a detectable level with the biomarker of the invention but not a peptide containing an unrelated sequence or a sequence of a different polypeptide. Binding properties can be assessed using an ELISA that can be readily performed by those skilled in the art. The binding agent may be a ribosome, RNA or DNA molecule or polypeptide with or without a peptide component. The binding agent may be a polypeptide comprising a polypeptide biomarker sequence, a peptide variant thereof, or a non-peptide mimetic of such a sequence. Aptamers include DNA or RNA molecules that bind nucleic acids and proteins. Aptamers that bind the markers of the present invention can be generated using conventional techniques without undue experimentation. [See, for example, the following publications describing in vitro selection of aptamers: Klug et al., Mol. Biol. Reports 20:97-107 (1994); Wallis et al., Chem. Biol. 2:543-552 (1995); Ellington, Curr. Biol. 4:427-429 (1994); Lato et al., Chem. Biol. 2:291-303 (1995); Conrad et al., Mol. Div. 1:69-78 (1995); and Uphoff et al., Curr. Opin . Struct. Biol. 6:281-287 (1996)]. Antibodies useful in the present invention include, but are not limited to, synthetic antibodies, monoclonal antibodies, polyclonal antibodies, recombinant antibodies, antibody fragments (eg, Fab, Fab', F(ab')2), dAbs (domain antibodies; see Ward et al. , 1989, Nature, 341:544-546), antibody heavy chain, intrabody, humanized antibody, human antibody, antibody light chain, single-chain Fv (scFv) (eg, including monospecific, bispecific, etc.) , anti-idiotypic (ant-Id) antibodies, proteins comprising antibody moieties, chimeric antibodies (eg, antibodies containing the binding specificity of a murine antibody but where the remainder is of human origin), derivatives such as enzyme conjugates or labeled Derivatives, diabodies, linear antibodies, disulfide-linked Fvs (sdFv), multispecific antibodies (e.g. bispecific antibodies), epitope binding fragments of any of the foregoing, and antigen recognition sites comprising the desired specificity any other modified configuration of the immunoglobulin molecule. Antibodies include antibodies of any class (e.g., IgA, IgD, IgE, IgG, IgM, and IgY), any class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or any subclass (e.g., IgG2a and IgG2b), and antibodies do not have to be is any particular type, class or subclass. In certain embodiments of the invention, the antibody is an IgG antibody or a class or subclass thereof. Antibodies can be from any animal source, including birds and mammals (eg, human, murine, donkey, sheep, rabbit, goat, guinea pig, camel, horse, or chicken). For example, antibodies for use in the present invention are commercially available from, for example, Invitrogen (Cat. Nos. PA5-83253, PA5-40331, PA5-40330, etc.), Abcam (eg, an antibody with Cat. No. ab224149), and the like. Alternatively, the antibodies can be prepared by recombinant methods well known in the art. In some embodiments, the antibody is a monoclonal antibody. For the preparation of monoclonal antibodies see, eg, Kohler et al. (1975) Nature 256:495-497; Kozbor et al. (1985) J. Immunol Methods 81:31-42; Cote et al. (1983) Proc Natl Acad Sci 80:2026-2030 and Cole et al. (1984) Mol Cell Biol 62: 109-120. The kits of the present invention can be prepared by conventional methods in the art. The kits may comprise materials or reagents (including reagents for detecting the RBM10 gene or its mRNA or its encoded protein or protein fragment) for use in carrying out the methods of the invention. Kits may include storage reaction reagents (eg, primers, dNTPs, enzymes, etc. in suitable containers) and/or support materials (eg, buffers, instructions for performing assays, etc.). For example, a kit can include one or more containers (eg, boxes) containing the corresponding reagents and/or support materials. Such contents may be delivered to the intended recipient together or separately. As an example, a kit can contain reagents, buffers, and instructions for use in detecting the RBM10 gene or its mRNA or its encoded protein or protein fragment. Kits may also contain polymerases and dTNPs, among others. Kits can also contain internal standards for quality control, positive and negative controls, etc. The kit may also contain reagents for preparing nucleic acid, eg, DNA, from the sample. The above examples should not be construed as limiting the kits and their contents applicable to the present invention. A microarray refers to a solid support with a flat surface having an array of nucleic acids, each member of the array comprising identical copies of oligonucleotides or polynucleotides immobilized on spatially defined regions or sites, The regions or bitpoints do not overlap with regions or bitpoints of other members in the array; that is, the regions or bitpoints are spatially discrete. Furthermore, a spatially defined hybridization site may be "addressable" in that its location and the identity of its immobilized oligonucleotide are known or predetermined (eg, known or predetermined prior to its use) of). Typically the oligonucleotide or polynucleotide is single stranded and is usually covalently attached to a solid support from either the 5'- or 3'-end. The density of nucleic acids containing non-overlapping regions in the microarray is typically greater than 100/cm 2 , more preferably greater than 1000/cm 2 . Microarray technology is disclosed, for example, in the following references: Microarrays: A Practical Approach, edited by Schena (IRL Press, Oxford, 2000); Southern, Current Opin. Chem. Biol., 2:404-410, 1998, the entire contents of which are via Reference is incorporated herein. The invention discloses the use of the RBM10 gene, and those skilled in the art can learn from the content of this article and appropriately improve the process parameters to achieve. It should be particularly pointed out that all similar substitutions and modifications are obvious to those skilled in the art, and they are deemed to be included in the present invention. The uses described in the present invention have been described through the preferred embodiments, and it is obvious that relevant persons can make changes or appropriate changes and combinations of the uses described herein without departing from the content, spirit and scope of the present invention, so as to realize and apply the technology of the present invention . The use of the RBM10 gene provided by the present invention will be further described below. Example 1 : Cioronib Monotherapy for Relapsed and Refractory Ovarian Cancer Phase Ib Clinical Trial Test Drug: Cioroni Capsules, Specifications: 5mg, 25mg. Produced by Shenzhen Microchip Biotechnology Co., Ltd. Dosing regimen: Cioroni capsules were administered at 50 mg/day, QD (no adjustment for body weight or body surface area). Take each morning on an empty stomach, with water, and swallow the capsule whole. Continuous administration for 28 days is one treatment cycle, and there is no interval between each treatment cycle. Number of cases: 25 patients were enrolled, of which 23 patients were evaluated for efficacy. Inclusion criteria: 1. Age ≥18 years old, ≤70 years old, female; 2. Histologically diagnosed epithelial ovarian cancer, fallopian tube cancer or primary peritoneal cancer; 3. Subjects must have received platinum-based chemotherapy regimens , that is, a) platinum-resistant patients (disease progression or recurrence ≤6 months after the end of platinum-containing chemotherapy regimens), should have received at least 2 different chemotherapy regimens after disease progression or recurrence; b) if platinum-sensitive patients ( Disease progression or recurrence > 6 months after the end of platinum-based chemotherapy regimen), disease progression or recurrence after receiving at least 2 chemotherapy regimens, or the subject refuses to receive further chemotherapy; 4. According to RECIST1.1 criteria, at least A measurable target lesion; 5. ECOG physical fitness score of 0-1; 6. The interval from the end of the previous chemotherapy, radiotherapy, targeted therapy, immunotherapy or study drug treatment should be more than 4 weeks, if the chemotherapy regimen includes mitosis 7. The main organ functions meet the following criteria: Blood routine: absolute value of neutrophils ≥ 1.5×10 9 /L, platelets ≥ 90×10 9 /L, hemoglobin ≥ 90g/L ; Blood biochemistry: total bilirubin ≤ 1.5 times the upper limit of the reference range (ULN), AST, ALT ≤ 2 times ULN (liver metastases: ≤ 5 times ULN); serum creatinine ≤ 1.5 times ULN; coagulation function: thrombin Original time-international normalized ratio (PT-INR) ≤ 1.5 times ULN. 8. Expected survival time ≥ 3 months; 9. Voluntarily signed a written informed consent. Treatment plan: test subjects take cioroni capsules 50 mg orally once a day, every 28 days as a treatment cycle, and there is no stopping interval between treatment cycles. Throughout the trial, all subjects continued treatment until any of the following occurred (whichever occurred first): disease progression, intolerable toxicity, death, withdrawal of informed consent, or loss to follow-up. Efficacy evaluation: According to RECIST1.1 criteria, the evaluation was performed at the baseline and the 4th week after treatment, and repeated every 8 weeks until disease progression. Tumor imaging examinations include CT or MRI of the neck, chest, whole abdomen, and pelvis. Examinations of other parts are performed according to clinical indications and when necessary. The same techniques and methods should be used for baseline and follow-up assessment of lesions. Safety assessment: including physical examination, vital signs, ECOG performance score, blood routine, urine routine, 12-lead ECG, blood biochemistry, electrolytes, coagulation function, cardiac enzymes, troponin, TSH, FT3, FT4, amylase, Echocardiography, 24-hour urine protein quantification (if necessary), adverse events. Biomarker study: 10 ml of peripheral blood was taken before the subjects took cioroni for the first time and when the disease progressed, and the gene sequence of plasma cell-free tumor DNA (ctDNA) and leukocyte extracted DNA (control) were detected, including a total of 548 A tumor-related gene, and the test results include gene mutation and copy number abnormality. Clinical Trial Results: Of the 25 subjects, 2 had no post-baseline efficacy assessment results, resulting in 23 evaluable cases. Among the 23 evaluable subjects, the best response during the trial was: 2 (8.7%) PR (of which 1 was confirmed at follow-up assessment), 14 (60.9%) SD, 7 (30.4%) PD , the confirmed ORR was 4.3% and the unconfirmed ORR was 8.7%. Of the 23 evaluable subjects, 3 were platinum-sensitive and 20 were platinum-resistant. Both PR subjects were platinum-resistant subjects, and the confirmed response was 1 (5.0%). The results show that cioronib monotherapy is effective in ovarian cancer. A concomitant study of efficacy-related biomarkers was performed on 548 tumor-related genes for the detection and analysis of cell-free tumor DNA (ctDNA) in the plasma of the evaluated patients. The detection results selected all genes with a mutation rate of more than 0.4%, and took the patient's progression-free survival (PFS) as the efficacy indicator to analyze the correlation between tumor-related gene abnormalities and the efficacy of cioronib. The results showed that among 548 tumor-related genes, a gene mutation that was significantly correlated with the efficacy of cioronib was found, which was the RBM10 gene. The median PFS was 232 days in 7 of the 23 evaluable subjects with RBM10 mutations (30.4%; these mutations included nucleotide additions, deletions and substitutions, and copy number changes) and 16 with RBM10 wild-type (69.6%). The median PFS of ) was 106 days, and there was a significant difference between the two groups, suggesting that the RBM10 gene is associated with the efficacy of cioronib in the treatment of ovarian cancer and can be a potential biomarker for evaluating the efficacy. See Table 1 and Figure 1 for the results. The above are only the preferred embodiments of the present invention. It should be pointed out that for those skilled in the art, without departing from the principles of the present invention, several improvements and modifications can be made. It should be regarded as the protection scope of the present invention.
