nini的BLOGISH

Dear Jennifer,


Nice to talk with you, I am Kung Hsiu-Ni. I heard from Prof. Chi that you are looking
for the T90 fellowship, and that is really of great interest to me. I am eager to apply
this job in order to gain more experiences in an exporter's Lab.

I have obtained my PhD in the School of Medicine, National Taiwan University (NTU),
where I am also a post-doctoral fellow. The NTU Medical School is the best medical school
in Taiwan. In my past 8 year experience, I focused on the biological and molecular
mechanisms of compounds, which are used as good chemotherapeutic agents on carcinoma
cells, working on different cells and animals. I am interested in some of the
hemoglobinopathies which are quite common in Asia. With the work and training experiences in Taiwan, I am confident that I can bring to the position the best I can offer.


The attached files are application materials and research
proposal. I hope my application will be appropriate for the T90
fellowship.

Thank you very much for your assistance.

Yours faithfully,

Kung Hsiu-Ni
post-doctoral fellow
Department of Anatomy and Cell Biology
College of Medicine
National Taiwan University
1-1, Jen-Ai Road,
Taipei, 100, TAIWAN
e-mail: d92446007@ntu.edu.tw
Tel: +886-2-23123456-88621
Fax: +886-2-23915292

Research experience
In my graduate study and postdoctoral training, I have been mainly working on the mechanistic understanding on how chemicals extracted from the traditional herbal medicines affect various cellular properties and biological processes. I focused on two compounds, β-lapachone (popularly used in South America) and tanshinone IIA (widely used in China). Both agents have been regarded as good chemotherapeutical agents against certain malignant carcinoma cells. We have performed various experiments to gain mechanistic understandings of their therapeutic benefits. In our past studies, we demonstrated that β-lapachone induced apoptosis of human prostate carcinoma (HPC) cells through the induction of endoplasmic reticulum (ER) stress, as shown by increase of intracellular calcium levels and induction of GRP-78 and GADD-153 proteins. Such β-lapachone-induced ER stress is accompanied by cleavage of procaspase-12 and phosphorylation of p38, ERK, and JNK, followed by activation of the executioner caspases, caspase-7 and calpain (Lien et al., 2008), suggesting that the endoplasmic reticulum is a target of β-lapachone for its therapeutic benefit. In a complimentary study, we also focused on the molecular mechanism of the anti-angiogenic effect of β-lapachone on causing the cell death of endothelial cell. We have demonstrated that β-lapachone induces endothelial cell death through the decrease in intracellular cGMP level and the mitochondria membrane potential, increase in cytoplasmic calcium and the resulting activation of calpain. We have further shown that nitric oxide (NO) can significantly attenuate the toxicity of β-lapachone and rescue human endothelial cells from apoptosis (Kung et al., 2007). Based on the utility of lapacho tree for improving the wound healing, I further investigated the effects of therapeutic dose of β-lapachone on several processes relevant for wound healing, including cell proliferation, cell migration, and in vitro and in vivo wound healing. We found that β-lapachone not only increases the proliferation and migration of cultured epithelial/endothelial cells but also improves the in vitro wound healing model using culture cells and in vivo animal model (Kung et al., 2008).
In the study of tanshinone IIA, we have demonstrated it triggers the death of human endothelial cell through activation of NQO1, which induces an increase of intracellular calcium and subsequently triggers the release of cytochrome c and induces the loss of mitochondrial membrane potential, thus resulting in the subsequent activation of caspases (Yang et al., 2005).
To further investigate the biological effects of β-lapachone on a global scale, we used DNA microarrays to test the gene expression at various dose of β-lapachone. Preliminary analysis of these gene expression data indicated that there are significant changes in the expression levels of several genes involving processes of endothelial cell death. These genes include osteopontin (ONP, SPP1), Caveolin-1 (Cav-1) and DEK. We are conducting functional studies to identify which of these candidate genes are responsible for the decision between proliferation and death of endothelial cell through the downregulation or overexpression using lentiviral systems.
In addition to the abovementioned research experience, I am also experienced in the animal handling, histological and molecular techniques. Moreover, I also help in writing part of research projects for applying grants and in the lab management. These experiences will also enhance my ability to contribute to the academic environments of Duke Medical Centers.

References

1. Kung, H.N., C.L.Chien, G.Y.Chau, M.J.Don, K.S.Lu, and Y.P.Chau. 2007. Involvement of NO/cGMP signaling in the apoptotic and anti-angiogenic effects of beta-lapachone on endothelial cells in vitro. J. Cell Physiol 211:522-532.
2. Kung, H.N., M.J.Yang, C.F.Chang, Y.P.Chau, and K.S.Lu. 2008. In vitro and in vivo wound healing-promoting activities of {beta}-lapachone. Am. J. Physiol Cell Physiol 295:C931-C943.
3. Lien, Y.C., H.N.Kung, K.S.Lu, C.J.Jeng, and Y.P.Chau. 2008. Involvement of endoplasmic reticulum stress and activation of MAP kinases in beta-lapachone-induced human prostate cancer cell apoptosis. Histol. Histopathol. 23:1299-1308.
4. Yang, L.J., C.J.Jeng, H.N.Kung, C.C.Chang, A.G.Wang, G.Y.Chau, M.J.Don, and Y.P.Chau. 2005. Tanshinone IIA isolated from Salvia miltiorrhiza elicits the cell death of human endothelial cells. J. Biomed. Sci. 12:347-361.

[下午 09:08:25] jentsan_chi-duke計老師 說 : please do not worry
[下午 09:08:38] jentsan_chi-duke計老師 說 : people are not mean
[下午 09:08:51] jentsan_chi-duke計老師 說 : they just want to chat about science
[下午 09:09:10] jentsan_chi-duke計老師 說 : pretent you are talking to people about your projects and your future research direction
[下午 09:09:53] [秀妮] 說 : I see
[下午 09:10:20] [秀妮] 說 : I will take it easier
[下午 09:10:26] [秀妮] 說 : thank you
[下午 09:10:35] jentsan_chi-duke計老師 說 : just get used to talking about your science
[下午 09:10:47] jentsan_chi-duke計老師 說 : communication is big in US
[下午 09:10:52] jentsan_chi-duke計老師 說 : writing grant
[下午 09:10:54] jentsan_chi-duke計老師 說 : writing paper
[下午 09:11:01] jentsan_chi-duke計老師 說 : giving talk is all communication

Q1: Why do you want to get this project in Duke?
A: Thalassemia is a serious problem in Asia, and thalassemia major is a huge burden to not only their families but also the government. I want to improve our understanding of the pathological mechanisms and microRNA regulation in thalassemia gene disorder and the malaria-infection resistance. The techniques of microRNA microarray and malaria infection on RBC are well established in Prof. Chi's Lab. I want to join Prof. Chi's lab to learn all the techniques and thoughts to find out greater molecular therapy for thalassemias. Prof. Chi recommended me to apply this fellowship to get the great opportunity to work in Duke. After few years of work, I wish I can develop better therapeutic strategies to improve the clinical outcomes of this disease and help thalassemias in Asia when I get back to Taiwan.

Q2: Where can you get samples?
A: Thalassemia is a common genetic blood disease around the world, especially in Asia. We have to compare thalassemias from different regions in Asia and America. We can get samples from Duke thalassemia therapy core, Taiwan and Thailand. We cooperate with Dr. Chang in National Taiwan University Hospital and Dr. Telen in Thailand. They will collect blood samples from parents of patients with thalassemia-major, since the parents are definitely thalassemia carriers. And they will send samples to Prof. Chi's Lab to do further analysis.

Q3: What is the plan about your husband and child?
A: My husband is a computer engineer working in Motorola in Taiwan, and he may go to America few months than me after finishing his job in Taiwan and finding a job in America. I have a baby boy; he will stay in Taiwan at first. As if my husband can find a job there, maybe we will take him to America.

Q4: You have not done this sort of job (work) before. How will you cope/succeed?
A: I have 8 years experience in science, and I try to solve problems with different thought and open mind. Although I am not the expert in regulation of microRNA on thalassemia, I will do my best to learn the new knowledge and to face all challenges.

Q5: Do you prefer working with others or by yourself?
A: It is fine to me either I work alone or cooperate with others. I am an independent person always thinking plans to solve problems and I also go along well with others when we work together. When I have problems that I can not figure out, I search references and discuss with people to find out the best resolution.

Q6: What is your major weakness?
A: I am a careful person always searching references and discussing with other people before making plans or decisions. So I need a period of time to prepare all things needed. Then I will follow the plan until it is finished. So my major weakness is time concerns of period of time.

Q7: What are your own special abilities?
A: I have strong sense of confidence with a positive attitude. I not only work independently with great enthusiasm but also go along well with everybody. I am responsible for my own job and always have clear sense of purpose. I firmly believe that I can succeed only with a constructive attitude and an open mind.

Q8: What is your career plan?
A: I will work in America for few years, and then get back to Taiwan to get a job in medical school and establish my own lab to work on thalassemia continuously. I wish I can contribute to thalassemia in Asia, especially in Taiwan.

Name: Kung Hsiu-Ni

Spouse: Lee, Ping-Hsun (1979/02/26)

Child: Lee, Yi-Hsung (2007/06/25)

Date of Birth: 7th January 1980

Residence: Taipei

Nationality: Taiwan

Telephone: [O] +886-2-23123456-88621
[H] +886-2-29861458

FAX: [O] +886-2-23915292

Email: d92446007@ntu.edu.tw

Address: [O] Department of Anatomy and Cell Biology, College of Medicine, National Taiwan University，1-1，Jen-Ai Road, Taipei , TAIWAN

Education: 2008 – 2003 National Taiwan University, Taiwan, Ph.D
Anatomy and Cell biology

2003 – 2001 National Yang-Ming University, Taiwan, MSc
Anatomy and Cell biology
(A course)

2001 – 1997 National Chung-Hsing University, Taiwan, BSc Zoology

Professional specialty:
Anatomy
Cell biology
Zoology
Cancer
Embryology
Histology
Biological Electron microscopy
Molecular biology

Working experience:
2008/08-- Postdoctoral position in National Taiwan University

Publications:

Papers—

1. Yang, L.J., C.J.Jeng, H.N.Kung, C.C.Chang, A.G.Wang, G.Y.Chau, M.J.Don, and Y.P.Chau. 2005. Tanshinone IIA isolated from Salvia miltiorrhiza elicits the cell death of human endothelial cells. J. Biomed. Sci. 12:347-361.
2. Kung, H.N., C.L.Chien, G.Y.Chau, M.J.Don, K.S.Lu, and Y.P.Chau. 2007. Involvement of NO/cGMP signaling in the apoptotic and anti-angiogenic effects of beta-lapachone on endothelial cells in vitro. J. Cell Physiol 211:522-532.
3. Kung, H.N., M.J.Yang, C.F.Chang, Y.P.Chau, and K.S.Lu. 2008. In vitro and in vivo wound healing-promoting activities of {beta}-lapachone. Am. J. Physiol Cell Physiol 295:C931-C943.
4. Lien, Y.C., H.N.Kung, K.S.Lu, C.J.Jeng, and Y.P.Chau. 2008. Involvement of endoplasmic reticulum stress and activation of MAP kinases in beta-lapachone-induced human prostate cancer cell apoptosis. Histol. Histopathol. 23:1299-1308.

Presentations—

1. Kung H. N., Chau Y. P., Lu K. S.
Microarray analysis of apoptotic cell death induced by beta-lapachone. September, 2005 APICA [OP-57]
2. Kung H. N., Lu K. S., Chau Y. P.
Nitric oxide attenuates the cytotoxicity of beta-lapachone on vascular endothelial cells. The 12th Joint Annual Conference of Biomedical Sciences, March 26, 2005 [P303]
3. Chau Y. P., Yang L. J., Kung H. N.
Tanshinone IIA elicits the cell death of human endothelial EAhy926 cells.
FEBS Journal 272, 2005 [N3-022P]
4. Hsiu-Ni Kung, Kuo-Shyan Lu, Yat-Pang Chau
MICROARRAY ANALYSIS OF APOPTOTIC CELL DEATH INDUCED BY BETA-LAPACHONE. The 13th Joint Annual Conference of Biomedical Sciences, March 19, 2006 [P425]
5. M.J. Yang , H.N. Kung , M.H. Pai, C.F Chang, K.M. Chang, K.S. Lu
BETA-LAPACHONE INCREASES THE PROLIFERATION RATE IN VITRO AND FACILITATES THE HEALING RATE OF SKIN WOUND IN VIVO. The 13th Joint Annual Conference of Biomedical Sciences, March 19, 2006 [P436]
6. H.N. Kung, C.F Chang, U.Z. Su, K.S. Lu
BETA-LAPACHONE PROMOTES WOUND HEALING. The 14th Joint Annual Conference of Biomedical Sciences, March 17, 2007[P785]
7. U.Z. Su, H.N. Kung, C.F Chang, K.S. Lu
MORPHOLOGY OF TASTE BUDS AND DISTRIBUTION OF GUSTATORY NERVE FIBERS IN THE CIRCUMVALLATE PAPILLA FROM MUTANT DYSTONIA (DT/DT) MICE AND DIABETIC (DB/DB) MICE. The 14th Joint Annual Conference of Biomedical Sciences, March 17, 2007[P788]

Personal Statement:

Nice to meet you, my name is Kung Hsiu-Ni. I was born as the eldest girl in a loving family in Chiayi County. I am an outgoing person with strong sense of self-discipline and interest in learning new things. While I was studying at university, I held the position as class representative, leader of general affairs in the class and in the student council within my department. I spared no effort while taking part in the activities held by the club.

I received a Bachelor’s degree in Zoology from National Chung-Hsing University, a M.Sc. in Anatomy and Cell biology from National Yang-Ming University and a Ph.D’s degree in Anatomy and cell biology from National Taiwan University. The master’s degree involved the anti-cancer effects of beta-lapachone and Tanshinone IIA, two natural compounds extracted from the traditional medicine and my Ph.D dissertation concerned the application of beta-lapachone on cell proliferation, cell migration and wound healing in normal or diabetic mice.

After graduation, I am now working in the Lab of Dr. Lu Kuo-Shyan in National Taiwan University. In this period, I work on the genes and proteins affected by beta-lapachone with microarray and some molecular and cell biology methods. I hope to find the most important effect and target genes of beta-lapachone on cells and animals.

I am an easygoing person, always there to help others. I especially enjoy acting like a counselor and providing advice when my friends encounter difficulties. I am a prudent person who would go through a thorough analysis process and collect related information before I make any kind of decision. I am an active leaning person; I face my life and career with a constructive attitude, for I deeply believe that only with an optimistic and open mind can I succeed. Pessimism and hesitation do not benefit my life. In school, I was interested in any section that I never know. I like to learn not only the knowledge for work but the other professional erudition, and I am ready to face all kinds of challenges with a can-do attitude. I hope that you could give me this opportunity to hold a position in your Lab. In the future, I will do my best in my task and study at every moment. Thank you very much for reviewing on my resume.

The thalassemias are a diverse group of genetic hemoglobinopathies characterized by absent or decreased production of normal hemoglobin, resulting in a microcytic anemia of varying degree. The thalassemias are classified according to which chain of the hemoglobin molecule is affected. In α-thalassemias, production of the α-globin chain is affected, while in β-thalassemias, production of the β-globin chain is affected. More than 90 types of mutation have been reported. The most common mutations of β-globin are point mutations, and six mutations in the hemoglobin genes account for over 90% of cases. More than 95% of the cases with β-globin gene deletion result in β-thalassemia major. In general, each population has a different group of mutations, consisting of a few very common ones and a variable number of rare ones. Thalassemia is the most common, inherited single gene disorder in the world, especially in Asia. In Taiwan, the prevalence of α-thalassemia trait is 4-5%, and β-thalassemia trait is 1-3%. The treatment of thalassemia major presents a heavy burden to all East Asian society. Although the Taiwan government initiated a National Screening Program in 1993 to reduce the number of patients with severe thalassemia, the thalassemia major is still a serious problem in Taiwan. Due to global migration patterns, there has also been an increase in the incidence of thalassemia in North America in the last ten years, primarily due to immigration from Southeast Asia.
Due to their effects on hemoglobins and anemia, all thalassemias affect the oxygen carrying capacity of the red blood cells and result in the morbidity and mortalities. The most severe forms of β-thalassemia major present within the first year of life with severe anemia and failure to thrive. Patients with severe thalassemia have to receive continuous blood transfusions and the resulting complications of iron overloading. This accumulation of excessive iron occurs in the major organs of the body, and is associated with tissue lesions, high risk for cardiovascular complications (e.g. iron-mediated cardiomyopathy), and other health problems.
The thalassemias have a distribution concomitant with areas where P. falciparum malaria is endemic. The α-thalassemias are concentrated in central and Southeast Asia, Malaysia, and Taiwan. The β-thalassemias are seen primarily in the areas surrounding Mediterranean Sea, Africa and Southeast Asia. Being a carrier of the disease has been shown to confer a degree of protection against malaria. This protection is manifested by their susceptibility to the less lethal species Plasmodium vivax malaria, while making the host RBC environment unsuitable for the lethal strain Plasmodium falciparum. It is commonly believed to that such resistance to malaria explains the high frequency of thalassemia in regions endemic for malaria since thalassemia provides survival advantages in the face of severe malaria infection. It is still unknown what cellular characteristics in the thalassemia erythrocytes are responsible for their relative resistance to Plasmodium falciparum infection. Understanding the molecular basis of this resistance will not only lead to a better understanding of malaria-erythrocyte interaction in thalassemia, but also may lead to novel approaches to prevention and treatment of malaria infection.
Recent study has shown the presence of microRNAs in mature erythrocytes. MicroRNAs are non-coding RNAs of 19-25 nt in size which mediate post-transcriptional regulation of target mRNAs through the formation of non-canonical base-pairing with the 3’UTR. MicroRNAs regulate a wide variety of biological processes (e.g. differentiation, apoptosis, oncogenic transformation). Several microRNAs have been also implicated in the process of erythropoiesis- miR-221/mir-222 and miR-24 are involved in downregulating proteins, including c-kit receptor and ALK4, those are responsible for erythrocyte differentiation. Several recent studies have investigated the role of microRNA during in vitro erythroid differentiation. Many important regulatory genes (e.g. GATA-1, EKLF) in erythropoiesis are also predicted to the mRNA targets for many microRNAs based on bioinformatics.
The global analysis of erythrocyte microRNA composition can inform the disease phenotypes of these sickle cell disease patients. Importantly, investigation on the malaria resistance of sickle cell disease has also implicated the dysregulated erythrocyte microRNA expression in sickle cell disease as genetic determinants of malaria resistance. This strategy can also be used to determine whether similar microRNA dysregulation may also play a role in the malaria resistance in thalassemia. My current proposal is to use global analysis of erythrocytes from patients with thalassemia alleles and disorders through the cooperation with Dr. Hsiu-Hao Chang in the Divisions of Hematology and Oncology of National Taiwan University Hospital as means to understand their disease phenotypes, clinical heterogeneity and their relative resistance to malaria parasites. As illustrated in the previous study of sickle cell diseases, the genomic study of microRNAs in mature erythrocyte provides an accessible and unique window to enhance our understanding of their regulatory roles in pathological states such as thalassemia and during P. Falciparum infection. To achieve this overall objective, we propose the following aims:

Specific Aim 1: Determine the erythrocyte microRNA gene expression of patients with thalassemia diseases

We will first recruit 20 thalassemia patients who are heterozygous and homozygous for -thalassemia from Taiwan and Duke. These patients will be consented to obtain blood samples for the isolation of mature erythrocytes for RNA purifications using the facilities (Leukocyte and reticulocyte depletion using Automac machines) available both in National Taiwan University and Duke Medical Center. The microRNA composition of erythrocytes from these patients will then be determined using microRNA microarrays in the Duke Medical Center. These patients’ samples will be hybridized together with reference RNAs from whole blood in the same manner as for sickle cell diseases and other anemia disorders. These expression data will be submitted into the Duke Microarray Database for further analysis. We will first compare the thalassemia microRNA expression with those of normal controls collected in these studies in an unsupervised analysis. Similarly, the microRNA expression of these samples will also be compared with those of sickle cell disease, PNH and other anemia disorders. Furthermore, we will perform supervised analysis of all the conditions to identify microRNAs whose expression is restricted to one or more than one anemia conditions. Finally, we will validate the differential expression of these microRNA using microRNA real-time PCR.

Specific Aim 2: Determine the role of dysregulated microRNA expression in the malaria resistance of thalassemia erythrocytes

In this aim, we will test whether the differentially expressed microRNAs in the thalassemia erythrocytes can contribute to their relative resistances to Plasmodium falciparum. We will first over-express several thalassemia-specific microRNAs in normal erythrocytes to determine their effects on the growth and proliferation of P. falciparum. The selection of these microRNAs will be based on the degree of dysregulation in thalassemia erythrocytes and previous known functions of these microRNAs. Afterwards, we will block these thalassemia-specific microRNAs to determine their effects on malaria growth in thalassemia erythrocytes. Finally, we will determine the 5’ end modification parasite target RNAs for the most functionally relevant thalassemia-specific microRNAs by using the in vivo capturing assays and malaria microarrays to define the potential relevant target RNAs in their anti-malaria activities.

The achievement of these aims using genomic approach to dissect thalassemia has the potential to have broad impacts in determining the role microRNA in the disease phenotypes and heterogeneity of thalassemia and malaria resistance to Plasmodium falciparum. Elucidating the role of microRNAs in regulating gene expression in the erythroid lineage will also improve our understanding of the pathological mechanisms in thalassemia and malaria diseases and allow development of better therapeutic strategies to improve the clinical outcomes of these hemoglobinopathies.