國立成功大學「邁向頂尖大學計畫」
延攬優秀人才「參與研究」計畫書
一、延攬理由（擬參與工作之重要性、其專長與計畫之配合程度、擬參與研究之工作內涵。
如為續聘申請案，請繳交前一聘期之工作報告。）
In this project it is essential to have a postdoctoral fellow participated in this project. In order to perform these experiments, I need a well-trained biologist to work with me to incorporate new experiments. Dr. Su has worked on bacterial virulence protein for her Ph.D. She has worked in my lab for seven years and expertise in protein expression, functional study, and protein-cell manipulation.
She is interested in pursuing research career. With her capacity for hard work and commitment for research career, she will be a good asset for my lab and for this study.
二、受聘人之工作內容。
In this study. Dr. Su will
1. optimize and design the di-specific (αVβ3 and αVβ5, αVβ3 and α5β1, or αVβ5 and α5β1), tri-specific (αVβ3, αVβ5, and α5β1), and multi-specific (αVβx or αVβx and α5β1) integrin drugs;
2. set up the new integrin-based cell adhesion assay, such as integrin αVβ3, αVβ1, α8β1, and α9β1, for the design of the specific integrin drug;
三、預期完成之工作項目及具體成果。
In our previous study we have successfully designed selective and potent integrins (αVβ3- and αVβx-specific disintegrins and integrin α5β1-specific FN-III 9-10 using Rho and FN-III as protein scaffolds. We anticipate that the development of di-specific (αVβ3 and αVβ5, or αVβ3 and α5β1), tri-specific (αVβ3, αVβ5, and α5β1), and multi-specific (αVβx or αVβx and α5β1) integrin drug will be done within three years. We also found that integrins αV, β1, and β5 were highly expressed in clinical HCC patients. In xenogragh animal model, we also found that integrins αVβx and α5β1 -specific disintegrin can reduce tumor growth. Therefore, we anticipate that one of the integrin-specific drugs will be a drug candidate for liver cancer treatment. We anticipate that the drug candidate will be in preclinical trial within three years and in clinical trial within five years. Three papers with SCI > 5 will be published, and one patent will be applied. We will make exclusive license and development agreement or co-develop our drug candidate with biotechnology company after finishing this study.
國立成功大學「邁向頂尖大學計畫」
延攬優秀人才工作報告表
NCKU’s “Aim for the Top University Project”
Work Report Form for Distinguished Scholars
□續聘continuation of employment ■離職resignation
受聘者姓名
Name of the Employee 蘇榆芳 □男 ■女
Male Female
聘 期 Period of Employment
From 107 年(y) 01 月(m) 01 日(d) To 107 年(y) 07 月(m) 31 日(d) 研究或教學或科技研發與
管理計畫名稱 The project title of research,
teaching, technology development and management
利用整合蛋白拮抗劑與天然物 發展抗癌藥物
計畫主持人
（申請單位主管）
Project Investigator (Head of Department/Center)
莊偉哲
補助延聘編號
Grant Number HUA107-3-7-051
一、 研究、教學、科技研發與管理工作全程經過概述。（由受聘人填寫）
Please summarize the entire research, teaching, or science and technology R&D and management work process (To be completed by the employee)
The research on the interaction between integrins and fibronectin (Fn) well facilitate the development of integrin antagonists. Especially, integrin α5β1 is the major receptor of fibronectin. RGD motif and PHSRN region are required for the interaction between fibronectin and integrin α5β1. However, RGD motif and PHSRN region are both required for the interaction between fibronectin and integrin αIIbβ3, though integrin αIIbβ3 is not the major receptor of fibronectin. As for integrin αvβ3, only RGD motif of
10Fn3 but not PHSRN region of 9Fn3 is required for integrin αvβ3-binding. Therefore, we proposed that
10Fn3 and 9,10Fn3 could be rationally designed to integrin αvβ3-specific and α5β1-specific antagonist, respectively. Engineering of sequence and disulfide bond can modulate activity and biophysical properties of proteins. So far, the work I have done as below:
Figure 1. Sequence alignment of wild type and mutants.
Residue L1408 was mutated to Proline as 9,10Fn3L1408P mutant. Disintegrin sequence
(RARGDNPD) is incorporated in residue 1491-1498. A disulfide bridge is introduced between residue 1490 and residue 1499. Identical amino acids among all four proteins were shown in dot.
Figure 2. The affinity to integrin α5β1 of 9,10Fn variants.
Affinity of mutants to integrin α5β1 was determined by inhibition of K562 cell adhesion to fibronectin.
Inhibition curves of 9,10Fn3L1408P (triangle), 9,10Fn3L1408P(RARGDNPD) (circle) and
9,10Fn3L1408P (CRARGDNPDC) (square) were shown. IC50 of cell adhesion to integrin α5β1 of 9,10Fn3L1408P, 9,10Fn3L1408P(RARGDNPD) and 9,10Fn3L1408P (CRARGDNPDC) were 245, 476 and 61 nM, respectively.
Figure 3. Selectivity to integrin α5β1 of 9,10Fn variants.
9,10Fn3L1408P (black), 9,10Fn3L1408P(RARGDNPD) (gray) and 9,10Fn3L1408P (CRARGDNPDC) (white) were tested the selectivity to different integrins (α5β1, αvβ3, and αIIbβ3) by cell adhesion inhibition assay. The results showed that the variants exhibited the selectivity to integrin α5β1 and no inhibition to CHO- β3 and CHO-αIIbβ3 cells adhesion to fibrinogen.
Figure 4. Sequence alignment of wild type and mutants.
Disintegrin sequence (PRGDMPD) was incorporated into residue 1492-1498. A disulfide bridge
proteins were shown in dot.
Figure 5. The affinity to integrin αvβ3 of 10Fn variants.
Affinity of mutants to integrin αvβ3 was determined by inhibition of αvβ3-overexpressed CHO cell adhesion to fibrinogen. Inhibition curves of 10Fn3 (square), 10Fn3(PRGDMPD) (triangle) and 10Fn3 (CPRGDMPDC) (circle) were shown. IC50 of cell adhesion to integrin αvβ3 of 10Fn3,
10Fn3(PRGDMPD) and 10Fn(CPRGDMPDC) were 4845, 610, and 99 nM, respectively.
Figure 6. Selectivity to integrin αvβ3 of 10Fn variants.
10Fn3 (black), 10Fn3(PRGDMPD) (gray) and 10Fn3(CPRGDMPDC) (white) were tested the selectivity to different integrins (α5β1, αvβ3, and αIIbβ3) by cell adhesion inhibition assay. The results showed that the variants exhibited the selectivity to integrin αVβ3 and no inhibition to α5β1 and CHO-αIIbβ3 cells adhesion to fibronectin or fibrinogen, respectively.
二、研究或教學或科技研發與管理成效評估（由計畫主持人或單位主管填寫）
Please evaluate the performance of research, teaching or science and technology R&D and management Work: (To be completed by Project Investigator or Head of Department/Center)
(1)是否達到延攬預期目標？
Has the expected goal of recruitment been achieved?
受聘人利用整合蛋白拮抗劑與天然物發展抗癌藥物之研究，達預期目標，未來可供實驗室繼 續進一步探討。
(2)研究或教學或科技研發與管理的方法、專業知識及進度如何？
What are the methods, professional knowledge, and progress of the research, teaching, or R&D and management work?
受聘人對於瞭解整合蛋白拮抗劑、發展拮抗劑與天然物發展抗癌藥物的機制與未來可行性之 探討有達到期望之進度，未來可繼續聘用執行研究目標。
(3)受延攬人之研究或教學或科技研發與管理成果對該計畫(或貴單位)助益如何？
How have the research, teaching, or R&D and management results of the employed person given benefit to the project (or your unit)?
受聘人在此期間對該計畫進行專業學術研究，對於其個人及受聘單位均有所助益。
(4)受延攬人於補助期間對貴單位或國內相關學術科技領域助益如何？
How has the employed person, during his or her term of employment, benefited your unit or the relevant domestic academic field?
受聘人在此聘期內的結果可供實驗室對於拮抗劑與發展抗癌藥物的領域繼續深入探討，其對 國內相關學術科技領域及未來醫學也有助益。
(5)具體工作績效或研究或教學或科技研發與管理成果：
Please describe the specific work performance, or the results of research, teaching, or R&D and management work:
受聘人在此期間對於整合蛋白拮抗劑發展為抗癌藥物的範疇有達成一定的成果，其工作結果
顯示利用9,10Fn3 作為骨架設計針對 α5β1 具有專一性之拮抗劑，成功的找到一突變體對於整合蛋
白 α5β1 具有高度親和力同時具有高度選擇性。之後將延續上述的成果做更進一步的優化此拮抗
劑之功能並探討其特性。綜合這些成果期望未來能發展出有效的抗癌藥物。
(6)是否續聘受聘人? Will you continue hiring the employed person? □續聘Yes ■不續聘No
※ 此報告表篇幅以三～四頁為原則。This report form should be limited to 3-4 pages in principle.
※ 此表格可上延攬優秀人才成果報告繳交說明網頁下載。
※ This report form can be downloaded in http://scholar.lib.ncku.edu.tw/explain/
