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Sep 16, 2020

Press Release: HKUMed researchers create a screening platform to discover new combination therapies for ovarian cancer and Parkinson’s disease

Press Release (2020-09-16):

HKUMed researchers create a screening platform to discover new combination therapies for ovarian cancer and Parkinson’s disease


Dr Alan Wong, Assistant Professor of the School of Biomedical Sciences, HKUMed (middle); and his team, Ms Becky Chan (1st left), Ms Cindy Tong (2nd left), Ms Yukki Wan (1st right), and Dr Nick Zhou (2nd right). 

The LKS Faculty of Medicine of The University of Hong Kong (HKUMed), in collaboration with School of Life Sciences of The Chinese University of Hong Kong, Faculty of Health Sciences of University of Macau, and the Ming Wai Lau Centre for Reparative Medicine of Karolinska Institute (Hong Kong node), have developed a new platform to systemically study the knockout effects of all possible two-way or three-way combinations in a given set of genes, with simple steps. By applying the platform to the druggable genes, which have existing drugs to inhibit their proteins, the team has successfully identified effective double or triple regimen combinations suppressing ovarian cancer cell growth and against Parkinson’s disease toxicity. Their work is now published in Cell Reports [link to the publication] and a patent application has been filed based on this work.



“No man is an island”, the same is also true for genes. Multiple genes co-operate to regulate almost all biological processes, including those driving diseases. Therefore, combination therapy, targeting multiple genetic elements, is developed, especially for complicated diseases, such as cancers and neurodegenerative diseases. Although it is promising, the traditional way to find the synergistic regimen combinations is always prohibitedly expensive and labour-intensive, which demands of testing all the possible combinations individually. Due to the limited manpower, the testing is hardly to be sufficient.



The team developed the CombiGEM-CRISPR v2.0 system to guide selection of drug combinations to be tested. Only a single set of gRNA oligos is required to build screening libraries, which drastically reduced the cost. Further, only one-pot sequencing of short barcode sequences is enough to guide the selection of effective two- and three- way combinations in this system. The merit of this platform is simplicity and extensibility. “Technically, there is only one step of cloning to make a one-way gRNA expression library to two-way, and another step to make it a three-way” stated by Dr Zhou, the first author of the work.   


Gene knockout could mimic the effects of the drug inhibitors on its protein. With the new platform in hand, the team decided to identify novel drug combinations for therapeutics of ovarian cancer, the 6th most common cancer among women in Hong Kong, and Parkinson’s disease, which has a prevalence of more than 1% in people over 60-year old in Hong Kong. Screening in the cancer cells, the research group found that knocking out DNMT1 + POLA1 + EGFR or ERBB2, as well as CDK4 + MAP2K1 + POLA1 in three-way combinations, and PARP1 + DNMT1 and CDK2 + FKBP1A in two-way combinations significantly suppress the cancer cell growth. Then it comes to the most exciting part. The researchers applied the drug inhibitors of each gene by the combinatorial way, the synergistic effects of suppressing the growth of multiple ovarian cancer cell lines are valid. In another screen to find the drug pairs against Parkinson’s disease toxicity, the researchers  identified double knockout of HSP90B1 + HDAC2 to be an effective combination for protecting cell death from induced PD-associated toxicity. The combination of matching drugs for these two gene products are also found to be effective and synergistic.


Significance of the study

With the new platform, researchers could rapidly identify effective combinations within three months from constructing complex gRNA libraries, estimated by the team lead Dr Alan Wong Siu-lun.  “It is hard to imagine how much effort and money would be spent to identify all those drug combinations by testing all the possibilities one by one. And we hope the drug combinations we discovered could eventually benefit those patients,” said Becky, the second author of the work. Indeed, PARP inhibitors are widely used as frontline therapeutics against ovarian cancer. One of our identified combination (PARP1 + DNMT1) has also been discovered by other groups for treating multiple cancers including ovarian cancer. Finding drug combinations is one important application of CombiGEM-CRISPR v2.0. “We hope our system could help identifying more unanticipated drug combinations that could benefit patients”, wished by the team.   


Previous study results

Dr Alan Wong Siu-lun has been developing various high throughput-screening technologies to systematically dissect the complex genetics underlying diseases including cancers and from there devising new therapeutic strategies. Previous work, published in Nature Biotechnology in 2015 and Proceedings of the National Academy of Sciences in 2016, focused on developing CombiGEM technology to screen genetic interactions, as the foundation of this current study. The team also published a review article in the Annual Review of Genetics in 2016 highlighting the potential of that technology in addressing practical challenges in biomedical research. Other than that, the team developed a new CombiSEAL technology published in Nature Methods in 2019, which extends the scope and impact of combinatorial genetics to engineer proteins for gene-editing applications.


About the research team

This study was led by Dr Alan Wong Siu-lun, Assistant Professor of the School of Biomedical Sciences, HKUMed. Dr Peng Zhou, Post-doctoral Fellow of the School of Biomedical Sciences, HKU, was the first author of this work. Ms Becky Chan Ka-ching, Ms Wan Yuk-kei, Ms Chaya Yuen Tsz-lo, Dr Gigi Choi Ching-gee, Ms Xinran Li, Ms Cindy S.W. Tong, Ms Sophia S.W. Zhong, Mr Jeiran Sun, and Mr Jien Vei Khaw (from HKUMed School of Biomedical Sciences and the HKU Biomedical Engineering Programme) were co-authors and they also performed the experiments. Other researchers were Prof Suet Yi Leung from Department of Pathology and Dr Lydia W.T. Cheung from School of Biomedical Sciences at HKUMed, Dr Zongli Zheng, Ms Yufan Bao, Ms Silvia Y.L. Mak, and Ms Maggie Z.Y. Chow from the Ming Wai Lau Centre for Reparative Medicine, Karolinska Institute in Hong Kong, Dr Wong Koon-ho and Dr Kaeling Tan from the Faculty of Health Sciences, University of Macau in Macau, and Prof CHAN Ho Yin Edwin from School of Life Sciences, CUHK in Hong Kong.



This work was supported by funds for Dr Alan Wong Siu-lun’s team including The University of Hong Kong Start-up and Internal funds, Hong Kong Research Grants Council (ECS-27105716, GRF-17102218, and TRS-T12-710/16-R), Innovation and Technology Commission (ITS/133/18), and Food and Health Bureau (HMRF-04151416). Other supports came from CUHK Gerald Choa Neuroscience Centre (7105306) (for Prof CHAN Ho Yin Edwin), the Research Services and Knowledge Transfer Office (MYRG2018-00017-FHS) and Faculty of Health Sciences (FHS collaboration and innovation grants) of the University of Macau (for Dr Wong Koon-ho’s team), and the Swedish Research Council (2016-02830) and the National Natural Science Foundation of China (81672098) (for Dr Zongli Zheng’s team). This project has been developed as part of the Ming Wai Lau Centre for Reparative Medicine Associate Member Programme, and Dr Alan Wong Siu-lun is a Ming Wai Lau Centre for Reparative Medicine Associate Member.


Media enquiries

School of Biomedical Sciences, HKUMed
Dr Wong Siu Lun Alan (Tel: 3917 9208 | Email:

School of Biomedical Sciences (General Office), HKUMed
Christina Chan (Tel: 3917 6334 | Email:


香港大學李嘉誠醫學院(HKUMed)與香港中文大學生命科學學院,澳門大學衛生科學學院以及卡羅琳醫學院劉鳴煒修復醫學中心(香港)合作,開發了一個可通過簡單步驟,有系統的研究給既定基因組合作雙向或三向組合敲取效應的新平台。 通過將該平台應用於可藥物基因,該團隊成功確定了有效的雙重或三聯療法組合,抑制卵巢癌細胞生長和抵抗柏金遜症的細胞毒性。 相關成果已於《Cell Reports》發表,並提交了專利申請 (按此瀏覽期刊文章)。



「沒有人是一個孤島」,基因亦是如此。 幾乎所有生物過程都是由多個基因共同調節,包括那些疾病相關基因調控網絡。 因此,醫療界開發了針對多靶點的聯合療法,特別是針對諸如癌症和神經退行性疾病等的複雜疾病。 盡管該單獨測試所有可能性組合的傳統療法前景廣闊,但卻受限於昂貴的成本以及大量的人力要求。鑑於有限的資源,傳統方法不足以找出大量有協同作用的藥物組合。



針對這一難題,該團隊開發了CombiGEM-CRISPR v2.0系統,去指導要測試的藥物組合選擇。 僅需一套gRNA寡核苷酸(oligos)即可構建高階多向篩選文庫,從而大大降低了成本。此外,針對於短條碼序列(barcode DNA sequence)的測序就足以識別有效的雙向和三向gRNA組合。 該平台的優點在於簡單以及可擴展性。 「技術層面上,僅需一個複製步驟就可將單向gRNA文庫變為雙向,再多一個步驟就是三向。」該研究的第一作者周鵬博士說。

基因敲除可以用來模擬藥物抑制劑對其蛋白的影響。借助新平台,研究小組決定探索可用於卵巢癌 ( 第6大威脅香港女性健康的癌症 ) 和柏金遜症 ( 六十歲以上香港長者的患病比率高至1% ) 治療的新型藥物組合。 在對癌細胞的篩選中,研究小組發現三向敲除DNMT1 + POLA1 + EGFR或ERBB2,以及CDK4 + MAP2K1 + POLA1,及兩向敲除PARP1 + DNMT1和CDK2 + FKBP1A可以顯著抑制癌細胞的生長。接下來研究人員通過以上組合方式應用了每個基因的藥物抑制劑,發現這些藥物組合可協同抑制多種卵巢癌細胞系的生長。 在另一個尋找抑制柏金遜症細胞毒性的成對藥物的篩選中,研究人員發現HSP90B1 + HDAC2的雙重敲除有效地保護了細胞免受柏金遜症相關毒性所引起的死亡。 此外,用於這兩種基因產物的匹配藥物組合亦有協同功效。

研究小組負責人黃兆麟博士估計,借助新平台,研究人員可以在三個月內完成從構建複雜的gRNA文庫確定有效的組合地任務。 「很難想象如果逐一測試所有可能,將花費多少精力和金錢才能找出所有這些藥物組合。 希望我們發現的藥物組合最終能夠使患者受益。」該工作的第二作者陳迦靜說。 實際上,PARP抑制劑已被廣泛用作抗卵巢癌的一線藥物。 其他小組也曾發現該研究中確定的組合之一(PARP1 + DNMT1),可用於治療包括卵巢癌在內的多種癌症。 尋找藥物組合是CombiGEM-CRISPR v2.0的一項重要應用。 「我們希望系統能夠幫助篩選出更多有益於患者預料不到的藥物組合」。



黃兆麟博士致力開發各種基於組合遺傳學的高通量篩選技術,用以剖析包括癌症在內等疾病的致病機理,從而制定新的治療策略。早前的研究工作開發了用於分析基因相互作用的CombiGEM技術。研究成果已於2015年在《自然 - 生物技術》和2016年在《美國國家科學院院刊》期刊上發表。此外,該團隊於2016年在國際期刊《遺傳學年度回顧》中發表了一篇評論文章,闡釋CombiGEM技術解決生物醫學研究的潛力和挑戰。 除此之外,該團隊還開發了一項新的CombiSEAL技術,發表於2019年《自然-方法》上,該技術將組合遺傳學的范圍和影響擴展到了蛋白質工程中,篩選出優化的基因編輯工具酶,Opti-SpCas9,以更好地進行基因編輯應用。



這項研究是由香港大學李嘉誠醫學院生物醫學學院黃兆麟助理教授領導。本研究的第一作者為香港大學醫學院博士後研究員周鵬博士。其他研究人員包括來自香港大學醫學院生物醫學學院和香港大學生物醫學工程項目的助理研究員陳迦靜,溫玉潔,博士生袁紫璐,博士後研究員蔡正姿博士,博士生李欣然,技術員唐思慧,本科生鍾碩華,孫介然和Jien Vei Khaw。其他研究人員包括香港大學病理學系的梁雪兒教授和生物醫學學院的張慧婷助理教授,香港中文大學生命科學學院的陳浩然教授,澳門大學健康科學學院的黃冠豪助理教授團隊,以及劉鳴煒復修醫學中心的鄭宗立助理教授團隊。



本研究主要獲得香港研究資助局(ECS-27105716,GRF-17102218和TRS-T12-710 / 16- R),創新與技術委員會(ITS / 133/18)和食品與衛生局(HMRF-04151416)。 其他支持來自香港中文大學的Gerald Choa神經科學中心(7105306),澳門大學研究服務和知識轉移辦公室(MYRG2018-00017-FHS)和健康科學學院(FHS合作與創新資助),瑞典研究委員會(2016-02830)和國家自然科學基金(81672098)。該項目是作為劉鳴煒修復醫學中心會員計劃的一部分而開發,黃兆麟博士是劉鳴煒修復醫學中心會員之一。



黃兆麟博士(電話:3917 9208 | 電郵

陳秀玲 Christina Chan (電話:3917 6334 | 電郵