Difference between revisions of "Team:CGU Taiwan/Human Practices Survey"

Survey

In our survey part, we have two main directions to look into the perception of experts and public. One is the judgements of our project received from the experts like doctors or scientists, especially in the field of immunology, infectious disease research, or anyone of concept-associated profession; the other is the investigation from the public to understand their acceptability of our product, "Leijuvant."

The practical way is that we came to visit doctors from the division of pediatric infectious diseases in Linkou Chang Gung Memorial Hospital and Taiwan Immunization Vision and Strategy, professors from Chang Gung University and National Health Research Institute, and scientists, business development division managers from vaccine-research-and-development companies like BOBC, Sanofi, Adimmune, and Medigen. And we put some issues to discuss with them, including the fields of project's research & development, clinical trials, business development, promotion, and regulations of laws or ethics, with the imaginary blueprint that our project is going to be commercialized in biotech industry (full question list). Here is the dialogue.

As for the examination of public perception, we used Google sheet as our questionnaire, distributing them to various population through the Internet. There are two main parts of questions we want to ask, including the apprehension of vaccine and their subjective point of views towards our concept.

We are very lucky to have reciprocal interaction with over 20 experts. Most of them agreed on our project, and gave us helpful feedbacks. Let's go through all the comments!

1. Results of our survey show that over 70 percent of participants appeal for the improvement of current adjuvants.

2.Results also show that nearly 60 percent of participants approve our product Leijuvant if it passes clinical trials.

3.Additionally, results demonstrate that virtually three fourths of people are more concerned about vaccine safety as compared to one fourth on vaccine efficacy. Very few people think about vaccine price and injection site. This survey outcome coincides with the comments from professional experts during our interview.

We visited many vaccine manufacturing companies and research institutes. The experts there gave us many suggestions and comments. Some key comments are quoted below. They are followed by our responses.

"Every vaccine product tends to be as simple as possible. If vaccine contains more compounds, more research will be needed. The product is then less likely to be launched due to increased uncertainty."

commented by Dr. Yang, R&D director of Adimmune Corporation

"A few of vaccines toward severe diseases are still ineffective. If a new vaccine has benefits outweighting its risks, such as side effects, the product may still be marketable."

said by Dr. Lee, Taiwan Immunization Vision and Strategy (TIVS)

It is known that Leishmania possesses a lipophosphoglycan coat over the cell surface. Lipophosphoglycan is a trigger for toll-like receptor II which initiates an innate immune response. There are also some uncertain modifications on Leishmania cell surface which may contribute to the specificity of Leishmania toward macrophage. In this project, we have proven the concept that Leishmania can serve as a potential vaccine adjuvant. In our future work, we will search for genes encoding Leishmania proteins that trigger immune response. The ultimate goal is to simplify our product and develop non-cellular adjuvant with same efficiency and fewer side effects. Moreover, we will explore the potential use of leijuvant in cancer or some incurable disease.

"The expense of R&D is the highest among others in developing a vaccine."

said by manager of Sanofi Pharmaceutical Company

In addition to R&D cost, culturing Leishmania will need large amount of medium, drug for selection, machines for electroporation and photo-inactivation. To address this issue, we will focus on severe diseases that still do not have effective therapy, like HIV, hepatitis C, malaria, and tuberculosis. As described above, developing a non-cellular system will also facilitate the process of scaling up. These two approaches should reduce manufacturing expense and attract big companies in investment.

"Even though there were only two to three cases of Leishmanaiasis in Taiwan decades ago, safety is still a concern both domestically and worldwide. If Leijuvant works effectively, the demands will increase considerably. You will then need to expand the scale of your factory and reduce the cost of land price simultaneously. As a way to reduce cost, constructing the factories in other countries means the safety issues will no longer remain domestic."

said by the manager of Sanofi Pharmaceutical Company

Leishmania depends on oxidative phosphorylation for survival, but is defective in the synthesis of heme required for electron transport complexes in mitochondrial respiration. This peculiar defect in biosynthesis is manifested as a nutritional requirement for heme in chemically defined medium. In nature, these parasitic protozoa must acquire heme exogenously from their hosts, such as sand fly, as a nutritional factor. Leishmania culture requires a certain osmolarity and combination of nutrients. Therefore, our transgenic Leishmania will not survive as soon as it leaves the medium. The outbreak of Leishmaniasis due to the leak live Leishmania from laboratory is nearly impossible. In addition, experiments involving live Leiahmania will be performed under P2 laboratory restrictions, such as wearing gloves and operating in a designated biosafety hood. One possible scenario of getting Leishmaniasis is when a laboratory worker accidently contacts high concentration of live Leishmania through an open wound and fails to initiate preventative measures immediately. Chance of this scenario becoming a reality is really low if people follow the proper guidelines. Because the vector sand flies do not exist in Taiwan, even an isolated case of Leishmaniasis won't spread to others.

What if sand fly suddenly appears due to climate change and the unlikely scenario does happen in our Leijuvant factory? Previous studies have shown that Leishmania expresses various 'virulence factors' in the sand fly, which may facilitate infection of the mammalian host. The alternative approach is to knock out the gene encoding virulence factors by genetic engineering to eradicate infection.

Since we provide another choice of adjuvant, establishing pharmaceutical quality should be a top priority. FDA in Taiwan regulates the industry via various standards, such as Current Good Manufacturing Practice (CGMPs) regulation for human pharmaceuticals. Many representatives in the industry we visited emphasized the important of CGMP regulation. We have searched the FDA website for potential regulations affecting our product. Before the clinical trials, we must go through the pre-clinical animal test. The approval of new drugs should be ethical during human efficacy studies. FDA will rely on evidence from animal studies to provide substantial evidence of effectiveness only when all of the following four criteria are met:

(1) There is a reasonably well-understood pathophysiological mechanism of the toxicity of the substance and its prevention or substantial reduction by the product.

There is a well-established step of photo-inactivation for our transgenic Leishmania in our Leijuvant manufacturing procedure. The dead Leishmania will not infect mice and dogs according to previous studies. However, we will still be cautious about the unknown or potential side effects of dead Leishmania.

(2) The effect is demonstrated in more than one animal species expected to react with a response predictive for humans, unless the effect is demonstrated in a single animal species that represents a sufficiently well-characterized animal model for predicting the response in humans.

In our project, we only use mouse as the model in evaluating immune response. If the mouse data show statistical significance, we will consider additional animal model testing before launching our leijuvant product. For example, rhesus macaque (Macaca mulatta) is a model well-known in developing several kinds of vaccine.

(3) The animal study endpoint is clearly related to the desired benefit in humans, generally the enhancement of survival or prevention of major morbidity.

Vaccination for certain infectious diseases, such as HIV, hepatitis C, malaria, and tuberculosis, is still not successful. It is mostly due to low level and non-protective responses from T and B cells.. We aim to facilitate both cellular and humoral immune responses by using Leijuvant as an alternative vaccine adjuvant.

(4) The data or information on the kinetics and pharmacodynamics of the product or other relevant data or information, in animals and humans, allows selection of an effective dose in humans.

In our experimental design, we tested the dose of dead Leshmania from 106 to 108 promastigotes in B6 mouse. However, the dose usage in human will require further assessment before clinical trials.

The last experts we went find was the doctors from the division of pediatric infectious diseases in Linkou Chang Gung Memorial Hospital. They had rapid comprehension and granted us for two reasons. First, they said we did a good job that we shared lots of vaccine-related knowledges on our Fan-Page, the general public will benefit from see those. With the overall unwidely acceptance of vaccination and with the advent of internet and increasing population of online users, doctors appeal for that professional knowledge should be promoted and exposed to people just like we did. Second, the dry-lab part of our project, McHug as an integrated website to predict the peptide presented on MHC, was praised significantly by doctors, because this can be used in all research of potential vaccines. That will truly benefit all researchers in vaccine development field.

We thanked doctors from left to right, Dr. Kuan Ying Huang, Dr. Cheng-Hsun Chiu, Dr. Chih-Jung Chen, Dr. Chen-Yen Kuo, Dr. Yhu-Chering Huang, and Dr. Yu-Chia Hsieh.

We iGEMers came to Medigen Vaccine Biologics Corporation to have a conference in Hsinchu Science Park located in Jubei. Despite long distance from CGU to Medigen, Allen and Ting Wan from Business Development and clinical Department intensely looked upon our project, considering it a serious case, listening to our presentation, and thereby discussing with us heartily. They suggested that we put emphasis on the motivation (application of tough, unsolved diseases), eminent advantages of Leishmania (dual functional of CD4 and CD8 T cell stimulation), solid accordance (to safety concern), acceptable but not-to-the-point answering skills (to the 100% inactivation concern), compliance with approach of Leishmania conjugation, and manifesting the significance and impact on society of human practice.

To sum up, we're really appreciating their goodwill. And we got to know another splendid biotech company with gorgeous interior design and outstanding faculties.

We headed south to National Health Research Institute in Chunan, introducing our project to Dr. Shih-Jen Liu (right 2), Dr. Hsin-Wei Chen (right 1), and An-Hsiang Shen (right 3). After they heard our story, they gave us helpful advices, including putting emphasis on the necessity of Leishmania as an adjuvant and better strategy of Leijuvant development. They said we could identify exact antigens that are specifically recognized by the antigen presenting cells, and thus we can fuse a specific protein of Leishmania with an antigen in our biobrick, letting it be expressed in the Leushmania, so that antigens could be detected much more rapidly and easily. Another suggestion is that we could design a construct with a tag next to the antigen gene. Then once the antigen is expressed, it will be signaled and transported to the surface, so that APC could engulf antigen carried on the surface of Leishmania faster, but it came out to be a conflict with our original thought, we reckoned tagged protein of antigen would be considered a broken one and get degraded; however, we have a great discussion on it.

We were sincerely thanking them for the professional feedbacks.

We were happy that Mr. Simon Kao (right 1), the vice president of business division, gave us a chance and invited Dr. Junie Ruey Chen (left 1) and Dr. Chin-Fen Yang (right 2) from the research and development division to listen to our project and give us scientific feedbacks. After interaction, we are no longer puzzled and under the apprehension that Leishmania could be considered a kind of expression system or a subunit vaccine, instead of an adjuvant. Their concern is that Leishmania itself contain so many antigens, and our Leishmania is absolutely a carrier. They even recommended us we should test the efficacy of anti-ovary antibody rather than just the expression in ELISA test of the In Vivo test. Finally, they complimented on us attending such a meaningful international competition, and encouraged us we should come to see the only outstanding professors in many of the Boston's universities. Take good advantage of our iGEM trip to the USA.

We were kindly thanking them for the professional opinions and inspiring encouragement.

We're very glad to have an academic exchange with TIVS. Director Doctor Ping-Ying, Lee and other very professional doctors gave us lots of responses, answered many questions regarding vaccine industry, and pointed the difficulty on the study and development of adjuvant and vaccine. Receiving these valuable opinions and feedback, we wish that Leishmania can be provided as a new approach in the field of adjuvant. Our transgenic Leishmania has the unique CD8 cytotoxic pathway to activate cell-mediated T cell, and most unsolved diseases are incurable mainly due to there are no certain amount of T cell could be induced. And also owing to high cost of Leishmania incubation, hence Leishmania as an adjuvant can be used like HPV, hepatitis C from HCV, malaria disease from plasmodium, and tuberculosis. This breakthrough idea help upgrade the value of Leishmania in the industry.

We were faithfully thanking them for the sharp and intense recommendation.

We greatly showed our respect and gratitude to Sanofi for their interaction with us as an event of human practice. We truly thanked general manager of Taiwan and Hong Kong region and Doctor Tsung-Chih Lai from vaccine research department for giving the chance. As a multinational corporation, we do believe they can offer us numerous amounts of valuable experience and advices from the globally biotech industry's point of view.

The questions asked by us can be majorly parted in 6 categories, including the opinions of our project, how to manufacture the vaccine and adjuvant, analysis of clinical trials, economic development and implementation, how to promote the vaccination, and regulations of laws or normal ethics. For further questions, please go into this website: http://prezi.com/xeyhwd-d-i5b/?utm_campaign=share&utm_medium=copy

Here comes the advices and feedbacks:

First: Our questionnaire needs more opinions of experts, so that statistics outcome can convince others. We still also can adopt the opinions of public but it depends on how we design the question in our questionnaire. Thus we may need to search for someone who are adept at designing questionnaire and statistics.

Second: Why was live attenuated vaccine acceptable in the very first beginning? They said from the commercial point of view, if epidemic outbreak is out of control and current techniques are not advanced enough to develop an inactivated vaccines, people have no choice but to accept relatively better live attenuated vaccines. Nowadays, there still remain live attenuated vaccines such as measles vaccines.

Third: Is adjvant essential in vaccine? They said if there are no adjuvants, it is okay to concentrate the antigens. There indeed exists the market demand, some adjuvants even being developed into cancer drugs.

Fourth: How big is the possibility that our Leijuvant gets approved by the clinical trials if the fact that inactivation of Leishmania up to 100% was proved? They said that preclinical trials, animal tests, 3-phrases clinical trials are long term process needing complicated examination and solid, very firm data to support. In accordance with statistics, the successful possibility of whole trials is 1 per million; and that of animal test is 1 per 10 thousand.

Fifth: How to do SWOT analysis, including strengths, weaknesses, opportunities, threats?

Sixth: If got approved by clinical trials, what are the probable challenge in foreseeable future? They said due to the difference between the scales of bench work and industry, it may require more costs on industrial production. And how adjuvants be used with vaccines together and what target antigen to choose are also the problems. What's worse, if looking on the dark side, people may have no faith in government and are unwilling to accept it. Our project aims to ultimately develop an adjuvant and used it with HA together to become a flu vaccine.

Seventh: From vaccine study to launch, how long will it take empirically? They said that time costs are extremely high with the average of 12 to 15 years. Outcome of vaccine research is always in the two extreme opposites, either no efficacy or severe adverse side effect. And the time needed will inevitably increase as people get to know more and laws get revised more and more strictly. The most rapid vaccine is the influenza vaccine with 6 to at most 24 months, because it needs to do quality control constantly to test its efficacy and potency; if something wrong happened to just one of the tests, all are going to be threw away and required to redo it again, and these are also the reasons of vaccines always being short of. Present 5 in 1 or 6 in 1 vaccines are only developed and successful in 2 companies of GSK7 and SANOFI; Japan has just proceeded to 3 in.

Eighth: In vaccine industry, what are the most adverse and sources-depleted places? They said research and development (R&D) is the most difficult part for it will bear the risk if crushed, and it will be just like throwing the money to the ocean. Most people know many sorts of vaccines should be developed, but only in vain ending up giving up. Biotech-associated industry is actually like the intensive capitalism game which the rich can play and take their chance.

We presented our project to Dr. Yu-sun Chang and Dr. Hsin-Pai Li when our experiments were just in the first progress. And both of the professors said the data should be displayed so that it can convince others. They also told us one data showing the biobrick in Leishmania was workable should at least be conducted.

We thanked both of them for the scientific comments.

The seminar this time with professors from the department of biomedical science had in fact 2 purposes. One was to introduce our project to professors in the first time, and the other was that we still needed a lot of financial help and certainly wish to receive their generous donation to meet our budgetary demands.

Then we indeed received small donations from professors. And they also gave us some comments. Our project reckoned Leishmania can be applied as a vaccine, but Dr. Sebastian D. Fugmann pointed if Leishmania expressed antigen are engulfed into antigen presenting cells, the complete structure of antigen will be ruined, then how could say that uncompleted antigen can be successfully recognized by B cells to be activated to the plasma cells? After our long days of searching information, we changed our original thought, alternatively calling it adjuvant. Another opinion from Dr. Shu-Yuan Yang focus on how to package our project, although we may not make the whole outcome through the experiments, we needed at least one firm datum to show and needed to realize that we should expressed in a charming, worthy, potential way.

We appreciate Dr. Sze-Cheng John Lo, Dr. Jin-Chung Chen, Dr. Hsin-Pai Li, Dr. Shu-Yuan Yang, and Dr. Sebastian D. Fugmann, Dr. Ming-Shi Shiao, Dr. Chao-Lan Yu for taking the time to share their thoughts with us. Their feedback are extremely valuable to the success of our project.

It was an occasion that graduates of master or PH.D would present the poster to the judges. Judges were professors from biomedical science graduate school at CGU, and we thought we could use this to spread our idea and get comments from them and also for fund-raising.

We were grateful to Dr. Mei-Jie Jou and Dr. Chia-Jung Yu for listening to our project, Dr. Jin-Chung Chen for granting us access to poster, and Miss Hsin-Yuan Wang for assisting decoration.