1. Full name: Pham Thi Hong Nhung                                           2. Sex: Female

3. Date of birth: 26/03/1988                                                          4. Place of birth: Hai Phong

5. Admission decision number: 4050/QĐ-KHTN-CTSV, dated on 19/9/2013  by Rector of VNU University of Science.

6. Changes in academic process:

- Extended time by decision No 1033/QD-DHKHTN and 597/QD-DHKHTN signed by Rector of  Rector of VNU University of Science.

- Decision No 3055/QD-DHKHTN, dated on 25/9/2019 by Rector of VNU University of Science that sending PhD. student back to office.

7. Official thesis title: Construction of recombinant neurokinin-1 receptor expression vector in CHO cells for application in drug discovery and development

8. Major: Genetics                                                                          9. Code: 94 20 101.21

10. Supervisors:            Assoc. Prof. Dr. Dinh Doan Long

                                    Assoc. Prof. Dr. Hoang Thi My Nhung

11. Summary of the new findings of the thesis

- A novel expression vector pSFV-KLEPT1.2 was constructed from the pSFV2 vector by joining a new DNA sequence that has additional affinity tails (Flag-tag and poly-Histidine-tag), thrombin cleavage site, and multiple cloning sites. The eGFP construct was cloned into the pSFV2 vector for expression kinetic studies. The human recombinant neurokinin-1 receptor gene (NK1R) was expressed according to the novel SFV expression system procedures. BHK-21 cells yielded about 50 x 1010 infectious recombinant particles/mL when cotransfected with recombinant and helper RNA and incubated for 42 hours. After 24 to 48 hours of infection with SFV, a large number of CHO cells expressing recombinant NK1R with full activity for molecular pharmacological studies.

- Comparison of U937, HeLa, and HEK-293 cell lines, natural CHO cell line did not express NK1R and is most suitable for human recombinant NK1R expression. In CHO cells, intracellular Ca2+ concentration increases when recombinant NK1R interacts with SP agonists and decreases when this receptor interacts with AP antagonists indicating that the receptor is fully functional.

- The molecular pharmacological model of NK1R for interaction with methanol extracts and some compounds from 10 Vietnamese medicinal plants was established. Extracts from Piper nigrum L, Stephania cambodica Gagnep and Styphnolobium japonicum (L.) Schott were found to exert inhibition on agonist-induced NK1R activity. The correlation between in vitro pharmacodynamic of the Stephania cambodica extracts with NK1R and rotundine in this extracts were initially found. Strong inhibition of NK1R was observed for extracts revealing the highest inhibitory potency for rotundine with IC50 of 0,88 µM. The NK1R antagonistic activities are relatively stable with P. nigrum extracts (IC50 between 5,7 and 60,5 g/mL) and S. japonicum extracts (IC50 between15,4 and 140,6 g/mL). NK1R is not the main molecular target in the antagonistic activity with methanol extracts of Cinchona officinalis L, Codonopsis javanica Blume, Eleusine indica L. Graerth., Eleusine indica L. Graerth., Orthosiphon stamineus Benth, Panax bipinnatifidus Seem, Panax stipuleanatus Tsai & Feng.

12. Practical applicability:

The CHO cell expressing human recombinant NK1R with full activity can be applied to screen for target compounds from Vietnamese medicinal plants. The research opens new prospects for herbal medicine development, standardization and modernization of natural origin products.

13. Further research directions:

- Extending the application of other target compounds screening on recombinant NK1R using the SFV expression system.

- Developing NK1R antagonists from rotudine, capsaicine and piperine (drug optimization, metabolic studies, preclinical studies,...).

- The expression of common mutant human NK1Rs, thereby assessing the impact of NK1R variants on drug response.

14. Thesis-related publications:

[1] Long Doan Dinh, Nhung Hong Thi Pham, Nhung My Thi Hoang, Cuong Trinh Tat, Van Hong Thi Nguyen, Lan Thuong Thi Vo, Huyen Thanh Pham, Kenneth Lundstrom (2015), “Expression of Human Neurokinin 1 Receptor in CHO Cells to  Evaluate Involvement with Vietnamese Medicinal Plant Extracts”, 12th Protein Expression in Animal Cells, pp.175.

[2] Long Doan Dinh, Nhung Hong Thi Pham, Nhung My Thi Hoang, Cuong Trinh Tat, Van Hong Thi Nguyen, Lan Thuong Thi Vo, Huyen Thanh Pham, Kenneth Lundstrom (2015), “Interaction of Vietnamese Medicinal Plant Extracts with Recombinantly Expressed Human Neurokinin 1 Receptor”, Journal of Planta Medica Letters 2(1), pp. e42 - e47.

[3] Patent: Process for the production of CHO cell expressing the human Neurokinin 1 receptor (hNK1R). Inventors: Dinh Doan Long, Vo Thi Thuong Lan, Hoang Thi My Nhung, Pham Thi Hong Nhung, Trinh Tat Cuong. Admission decision number 49722/QĐ-SHTT, dated on 25/07/2017 by National Office of Intellectual Property of Vietnam.

[4] Pham Thi Hong Nhung, Dinh Doan Long (2018), “Establishment of an in vitro Screening Model of Bioactive Compounds Using the Recombinant Central Nervous System Receptors”, VNU Journal of Science: Natural Sciences and Technology 34 (1): pp.105-110.