Team:DUT China A/Human Practices
CICCAN
Through amount of reference to successful Hp case constant
self-summarize, we finally concluded a 6-step set strategy to complete
Hp consistent with our project. We named them
:“Contact”，”illustrate”，”Conversation”，”Change”，”Advance”，”Nicole”.
Contact: In the process of our project in the researching term, we
accumulated our targets to somewhere we need to have a Hp, which we
call it "Valuable Library". We utilized this library to deal with many
problems in the way of our research. With the continuous accumulation
in the process of early stage and mid-term, we successfully encounter
lots of problems at various stages and never worry about possible
helpless situation. Of course the first step when we have matters is
to depend ourselves rather than just blindly seeking advice. The
library gives us more help in terms of our thoughts and vision.
Illustrate: A team should always clear their project progress,
problems and definitely their project. In terms of our project, we
focused our team's project plan and check our project in case any
likely problems. We had a list and it held up our problem that we have
not ruled out and though we always filled something new on the list,
the length of our list is shorter and shorter. Facing the problems
hard to solve, we will to shed light on the problem and check the
Library search may provide us with help of experts and the team.
Conversation: After we successfully handled ‘Illustrate’ and
‘Contact’, our human practices group would construct the
‘Conversation’ through every possible method. In the ‘Conversation’
section, we would explain our project and the problems we met, we
would record the communication process as well. We would take the next
step as soon as possible once we finished the ‘Conversation’ part.
Change:
We would reconsider and adjust our project according to our
conversation records, preventing our project come to a standstill.
Advance: Sometimes we were hesitated to decide among several
suggestions because we could not distinguish by the present situations
whether they were beneficial to our project, thus we had to give them
a try. Using different analyzing methods constantly during our
experiment, we could finally decide which suggestion was really
helpful and meaningful to improve our project.
Nicole:
As we reached to the ‘Nicole’ part, it indicates that our human
practices have successfully conducted. Although it might has some
drawbacks in this rapid project construction, this is the best case we
can come up with within the preparing period. Nowadays, it is not hard
to obtain the information we need. But it will be much easier and
handier to use the CICCAN system to get the information, including
human practices aim, conference records and project suggestions. By
using the CICCAN system, we can finally emend our project close to
perfect, avoid risks and successfully come to the ‘Nicole’ in time.
Why We
Come to iGEM
“If you have a dream, go get it.” That’s the response from our advisor
when we first pitch the idea of researching the mechanism of cancer.
So here we come with our accomplishments about CTCs and we sincerely
would like to receive more perspectives from other scholars in iGEM.
When it comes down to our drive of attending IGEM, our team have
focused on two aspects: learning and getting feedback. Throughout the
preparation for this conference, we have already learnt a lot about
experiments, communicating with Hp, how to minimize risks and how to
properly make choices. We truly realize that only learning in our own
circle is not enough to spawn a mature project. However, our team
never gives up easily and we still unswervingly believe that our
project will be able to contribute to the world eventually.
Throughout the entire project, we have encountered a lot of
challenges. Step by step, from the brainstorm of our theme, to the
decision of capturing CTCs, and to finally realize it in the lab, we
overcame all those barriers to finally reach our Wiki statement. We
continue to learn and break throughout the process. In the end, we
will bring our results to the conference, using this platform to
showcase our projects and accept feedback from our peers and other
scholars. Hopefully, we could carry the dream overseas to this amazing
continent and reply to ourselves with a satisfying result of our
dream.
Why
do we believe that our project will contribute to the community and
society?
Our project is not just for participating in this event. We certainly
have more expectations for this project to deliver more information
for future research. We firmly believe that the birth and use of any
kind of technology is accompanied by numerous experimental
verifications and sample analysis. Although our team may not be able
to think of a complete pathway to cure cancer in a short time, but our
project can provide a perfect experimental background for future
research. With our help, no team will encounter serious damage when
transferring, gene sequencing, doing experiments with CTC samples like
what Heipros met before. Therefore, we could ideally avoid the cost of
time and money on CTC-related research and, most importantly. we are
willing to help other iGEMer or research teams with full supports.
At the same time, we have brought new thinking to the community. Our
project technology focuses on the application of functional DNA, which
combines nanomaterials and biological fields to provide more options
in cross-disciplinary research. Plus, we plan to apply our aptamer
hydrogel initiation system to more fields, such as the precise capture
of rare cells. We hope to provide more choices for experimental
materials for more research in the future and assist researchers in
achieving their experimental goals of improving humanity.
How do
we use Hp to complete our work?
We decided to do tumor-related work during the project research phase,
but as we all know, the tumor field is wide-ranging. We also need to
select specific projects that can meet our laboratory conditions and
iGEM cycle to conduct research. We started with immunization and the
use of Car-T technology, but, after discussing with Professor Xu Jie,
we realized that our laboratory may not provide sufficient
experimental conditions. Therefore, we conducted a re-investigation of
the project. After our conversations with Professor Hong Rui from
Liaoning Provincial Cancer Hospital, we realized the deficiency of
current CTC capturing technology to meet the demand of medical and
scientific researchers. Therefore, we used the sample provided by the
hospital to develop a possible pathway to approach the problem.
We decided to use DNA materials to design a low-cost cell capture
strategy by first constructing a cell cage through aptamers and DNA
hydrogels, and then using quantum dots and magnetic techniques for
separation. However, after consulting from researcher Cuihuan Qing
from Chinese Academy of Sciences Shenzhen Institute of Advanced
Technology, we learned about DNA prospects for the future development
of the hydrogel materials. Its low-cost and highly controllable
characteristics really attracts us for learning more about it.
Thereafter, we visited Shanghai Pu Los company, which is a precise
medical core business. In the process of communication with other
science and technology team, They suggested that we could focus on DNA
hydrogel, and that we should abandon the magnetic separation
technology to reduce costs.
Furthermore, based on out communication with LanJing lab, we decided
to make a CTC capture kit. This decision also provides a reference for
our subsequent experimental system. In the course of the experiment,
we use our CICCAN program, which can be seen later, to avoid risks and
have contacted Akkaya and Liu Dongsheng in advance. Group
communication, solving our problems in the specific experimental
process. In parts verification process, we with the help of Professor
Wang Xinkui, successfully verified parts viability.
How do
we ensure our experiments are safe?
Team DUT China A attaches great importance to the safety and security
of both scientists and the environment. Our host lab, Zi Laboratories,
follows General requirements for laboratory biosafety in China.All the
members involved in the experiment were trained strictly before thanks
to their professional course, our professional experimental teachers
helped us to conduct systematic experimental safety training right
before the lab work start.We strictly obey the rules of the laboratory
practices and guidelines. Three cell lines including HeLa (Epithelial
cell adhesion molecule negative, EpCAM-negative), MCF-7
(EpCAM-positive) and A431 (EpCAM-positive) used I our project are
commercialized and widely-used in research, without any pathogen.The
final aim of our project is to detect circulating tumor cells with
human blood. However, we will not conduct any test with blood but take
white cells to simulate real blood environment and carry out
experiments under such conditions.
How do we connect our thoughts with education?
After we reach a long-term cooperation in education, we realized that
we are really willing to devote ourselves for the future of human
evolution in life sciences. However, the world needs talents from all
other fields besides biology. Thus, in the process of education, we
never discredit students to think from other perspectives. Moreover,
we are paying more attention to how to connect our thoughts and
education while teaching biological theory. We have been doing so in
the past few years. From the academic accomplishments obtained under
academic research, to our determination of unswervingly persisting our
original intention, we all convey these to the students in different
forms, affecting their inner hearts in subtle ways. We try to provide
the students with a broad scope of thinking based on the life
sciences. Even if the students prefer other fields, they will still be
able to apply what they have learnt here to impact their future
selves. Overall, we hope that through such cooperation, we can
contribute more to students’ personal development.
Details
HaploX
This company focuses on the tumor-targeting therapy. As a global leader in precision medicine, HaploX is devoted to integrating advanced techniques such as liquid biopsy, genome sequencing, artificial intelligence, and big data into cancer diagnosis.At the beginning of the project, we did not know which part of our design should be focused on, what specific requirements of the precision medicine should be met, what experiment parameter related to our project and design and whether the cost of our project can be afforded. We turned to HaploX for help and received their support. We got the help of HaploX in the following aspects.
1) what to focus on in our project
2) which hypotype of EpCAM we should use
3) display us with the latest strategy of capsuling the circulating tumor cells (CTCs)
4) offer the suggestion of abandoning the idea of isolating the CTCs by magnetic materials.
5) offer the response interval of the quantum dot for us and suggest to choose other visualization materials
After receiving the suggestions and guidance from HaploX, we modified our project’s main point to the DNA hydrogel and decided which hypotype of the EpCAM to use in the experiment. We also gave up the idea of using magnetic materials to extracting the CTCs, as well as the quantum dots.
In the end, we choose the rolling circle amplification (RCA, or R) to achieve the triggered-amplification attached to the capture. This strategy we took can decrease the cost greatly.
Bluepha Lab
Bluepha Lab belongs to Bluepha, with Synthetic Biology as the technology carrier, focus on cutting-edge life science education, also is the CCiC conference partner. Although our team is committed to saving humans from the pain of delayed cancer discovery, we are still unsure how the project will evolve and in what form provided us with ideas for making the kit. The Blupha Lab provided us with the idea of designing the kits and gave us the guidelines for us to follow according to the development of kit. With the help and guidance from Bluepha Lab, we quantified every reagent and were able to adjust the experimental system by ourselves. This activity assist us to improve the experimental system and become more rigorous about the lab work.
Professor Dongsheng Liu's group
Professor Dongsheng Liu's group located in the Department of Chemistry, Tsinghua University. They are interested in using biomolecules and their specific interaction in nanostructure fabrication and nanodevice design. We met the problem of incompatibility between the hydrogel system (pH7.9) and the cell system (pH7.4), as well as the experimental problem and cost problem in the process of cultivating the hydrogel. Professor Dongsheng Liu's group provide us with the following solutions. 1) They provided us with the solution of flow analyzer.
2) Suggest us to change to another buffer of the enzyme to overcome the problem of pH inappropriateness
3) Provide us with visual methods of pigment staining
4) Solved the problem of hydrogel caking
5) Provided us a simple method for hydrogel separation
Finally, we came to the solution of measuring the mechanical coefficient to detect whether the hydrogel was formed. By adjusting the buffer of the enzyme to realize the system integration, the visualization cost was reduced and the efficiency was improved, and the experimental problem at that time was solved.
Rui Tong
Rui TongDeputy director of the of Gynecologic Oncology, Liaoning Cancer Hospital
When we did not determine the subject direction of the project, we made a discussion and we decided to make a technical breakthrough in cancer. We hope that we can clarify our project direction through making communication with Doctor Tong. She point out that there is a problem of incompatibility between medical and research in CTC technology. Therefore, we intend to capture CTC cells with intact activity to provide research for research, and we make our project decisions.
Akkaya
Akkaya Master tutor of Dalian University of Technology
The protocol of using DNA polyacrylamide is not suitable to the ssDNA which is smaller than 100nt in the experiment. The resulting strip does not verify the formation of the ring template. Therefore, we asked her for help. With her analysis, the reason may be the inappropriateness of the ratio of Monomer acrylamide’s concentration and methylene diacrylamide’s in the polyacrylamide gel, which result in the failure. The advice from her is changing the concentration ratio to increase Cross linking rate and decrease gel size. So, we changed the former ratio,19:1,into the present one,29:1. Eventually, The resulting gel has such a high resolution, that it can be distinguished the ssDNA being circled
Huanqing Cui
Huanqing Cui The researcher of Shenzhen Institutes of Advanced Technology Chinese Academy of Science. Research on intelligent polymer materials, mainly includes responsive hydrogel, polymer brush, macromolecule self-assembly, etc.
He showed us the hydrogel components he works on, and the prospects of using hydrogels.
Wang xinkui
Wang xinkui Ph.D of engineering, associate professor and master supervisor of dalian university of technology and dalian institute of chemical physics.The main research direction is co2 catalytic conversion and nanocatalysis.
At the present stage, the application of short ssDNA adapter body mostly related to a kind of new materials (gold nanoparticles, colloidal particles), it is difficult for us to use the biological methods to verify the ssDNA and degree of the combination of the target.In order to verify the three parts from the summit of Heidelberg university in 2015, we seek the help of miss wang. She thought that we can use nano gold way to verify from the perspective of chemical. Eventually, we assisted teacher wang to verify parts of ssDNA by nano gold colorimetry. During the experiment after this, all parts of verification process was completed.
Huanan Wang
Huanan Wang Professor and vice President of the School of Life Science and Technology, Dalian University of Technology. His research interests include 1) self-repairing materials and biomedical applications, 2) stem cell microenvironment and stem cell therapy, 3) microfluidic technology and biomedical applications.
During the research and development of the project, there was a problem of system incompatibility. While we have not yet determined the cause, we suspected that it might be the incompatibility between hydrogel and cells.
By communicating with Mr. Wang, we received plenty of advices:
1) Hydrogel is safe and harmless to human body, and can protect cells
2) Provide laser particle size analyzer to assist us in completing physical properties characterization of hydrogel
After the communication with professor Wang, we continued to carry on the hydrogel experiment, looked for systemic problems in other aspects, and performed physical properties characterization of the hydrogel. In the experiment after this, we completed hydrogel characterization, visualization and quantification experiments and found that the pH system was incompatible and continued to improve in this respect.
Jie Xu
Professor of Fudan university, deputy editor of BMC Cancer and special editor of Frontiers in Pharmacology, the main developer of colorectal Cancer early screening kit. At the beginning of the project, we talked with professor Xu, who helped evaluate the experimental conditions, rejected the original car-t development plan and suggested that we should focus on cancer cells instead of immunology.
Qihui Shi
Qihui Shi Professor of Fuda University, researched on CTCs.
We talked to him during the development of the project, he suggested that we should use chip to capture, but we have to give up this scheme due to the high cost. At the same time, we understand the development status of CTC capture.