Human Practices

Contribution

Dr. Eid described from his experience the mental difficulties that children and youth experience following hair loss, and how this affects their self-image and coping with the disease. Dr. Eid stated that our project is a very important initiative, especially for adolescent children, for whom this is a sensitive issue. The child must make a difficult transition between the old, normal routine and a new routine that includes painful treatments, pressure on the family, and a new self-image. We asked him how he thought the product should be presented to patients and he emphasized that nothing should be guaranteed in this area. Dr. Eid gives lectures in nursing courses, in which he teaches how it is taboo to make promises to patients that cannot be kept. He explained to us that in such situations the patient clings to every possible promise and hope, and if he experiences disappointment from a medical/scientific factor, this may lead to a severe regression in treatment. Therefore, he recommends presenting the product in a modest way, which presents the pros alongside the cons and gives the patient room for control. Not to present the product as an absolute solution because there is no absolute truth in this field.

“Finding a product that will help prevent hair loss affects not only the hair but also the mental stability, the withdrawal and development, the daily functioning - this is an important tool that will allow the patient to feel a certain control over his life” J.G.Eid

Implementation

Following our conversation, we learned about the importance of our project and the necessity of such a product, and how to make it approachable for patients. By doing so we alternated our survey and rephrased it to be more suitable for patients and those in recovery. We updated our approach towards the end product and centered on its use as a chemotherapy-induced alopecia product.

One of the most common solutions for hair loss is wigs. In Israel, every cancer patient who loses his or her hair due to chemotherapy treatments is entitled to receive a wig for free.
But did you know that it takes at least 7 braids to make one wig?

As part of our desire to understand the problem and to contribute to the community, and to the cancer patient in particular, on June 15th we organized a hair donation event on the Technion campus. The event was held in collaboration with "Zicharon Menahem" association which works for cancer patients and their families.

With the help of three amazing hairdressers that came voluntarily, we managed to collect dozens of donations sent to the association.

We were touched to find out that the event reached not only undergraduate students but also included donors from graduate school, faculty members, and even people from outside the Technion who came just to donate their hair.

Each of the donors received a certificate of recognition for their donation and a small gift bag that we prepared especially for them. Beyond the hair donations, the event helped us raise awareness of the issue of hair loss as a result of chemotherapy, and how the existing solutions are only partial and there's a need for new solutions.

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After the hair donation event, we realized that help from a distance is not enough, and to understand the issue more deeply, we must meet with the patients themselves.

On August 28th, members of our group joined the charity association “Larger than Life” in organizing a pool party for families facing pediatric cancer. We believe that such events not only provide families with the opportunity to step away from the white walls of the hospital but can also serve as a connecting and supporting platform, where parents and their children meet people facing similar difficulties. During this day we met and heard stories of strong families, which gave us some insight into their life.

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To verify that our project is necessary and contributes to the world, with the advice of Dr. Eid we wrote a survey for cancer patients and those recovering. The survey consists of 23 short questions, regarding the feelings accompanying hair loss, the self-image following CIA, the opinions on the products that exist today, and the need for a new product.

It should be noted that because the survey touches on a sensitive topic and population, we had difficulty conveying it to the target audience and reaching a statistically sufficient number of people, which represents the entire community. Therefore, the following survey will represent the data collected from seven cancer patients.

The majority of respondents testified that hair loss due to chemotherapy has a great impact on the disease's difficulty and its significantly affects their mental state. Additionally, more than half of the respondents believe that the solutions that exist today for hair loss after chemotherapy are ineffective and affirm that if there was a product that prevents hair loss due to chemotherapy, they would want to use it.

Gathering all this information we were able to reach the relevant understanding regarding the need for our project, the problem we are trying to solve, and the cooperation required of us with the intended population

Click here to see our survey questions

Contribution

They explained that there are currently two main methods for extracting from plants: steaming and pressing. The methods vary from plant to plant and depend on the percentage of oil. From the discussion, it emerged that the extraction process is accompanied by depreciation, which harms the environment, and its disposal is accompanied by associated costs, which are added to the costs of the raw materials, and the ongoing growing costs of the plant. Hence, the existing extraction processes in the market are polluting, and expensive and don't necessarily address the problem of low efficiency that exists in plants such as Angelica gigas.

In addition, in extraction processes and in particular, in refining processes, it is difficult to maintain the medicinal properties of the extract. This makes, plant extracts not always suitable for such products. Another issue that came up in the conversation is the issue of standardization and how it is carried out in practice. In the conversation with Rachamim, we realized that standards are completely different between the food sector, cosmetics, and medicine.

Implementation

The coversationwith Shamna helped us understand the regulatory mechanisms that exist in the western world, and what we'll be required to do in the field of standardization for the synthetic decursin we will produce.

Yaki raised an important question - why are you trying to mimic nature? What's the point of trying to do something that nature already does?

From his questions, we understood how the project was perceived by the industry and stakeholders. Yaki, as an industrialist, was concerned about production costs. In his opinion, even a production that "imitates" nature will require additional costs in energy, raw materials, and people. For him, this is an economic and not a technological question. Following Yaki's concern as an optional stakeholder, we concluded that a feasibility model was required, which would be an essential component of the work plan and would justify the viability of the project. Read about it in the Proposed Implementation page.

Our meeting with Yaki Harel and Rahamim Eliyahu

Contribution

When we set out to biomanufacture decursin, we had a clear vision in mind (learn more in Proposed Implementation page): enabling the development of novel treatment strategies that make use of decursin. This meant thinking beyond manufacturing and led us to seek feedback regarding regulations, safety requirements, and screening processes. With the help of former iGEMer, Ricarina Rabinovitz, we set up a meeting with Pharmaseed which included a tour of their lab and a meeting with Dr. Itschak Lamensdorf who founded Pharamaseed in 2003, and today serves as a chairman and a consultant to the company.

Implementation

Pharmaseed shared from their experience on the path of new drug discovery. We quickly learned that biomanufacturing is only one side of the picture, and future strategies that include decursin would have to further research it's molecular mechanism. This was quite unexpected for us because while our team is on the biomanufacturing track, we understood that it’s impossible to divide science to different separate tracks, as synthetic biology integrates various disciplines. This helped us learn about the process of new drug approval, a core part of our proposed implementation. Furthermore, Pharmaseed offered their biological screening services as an opportunity to collaborate, while we can work on the biomanufacturing aspect.

Contribution

We were advised to use HPLC because every molecule involved in our pathway (decursin/decursinol, umbelliferone, 7-demethylsuberosin, and marmesin) is aromatic and will be suitable for separation using this technology.

Implementation

We took that information and conducted a literature survey to find existing protocols for using HPLC for measuring our metabolites. Once we had an initial direction, we worked with Fishman's lab back and forth to finalize HPLC program suitable for our samples. You can read more about our measuring process here in the Results page.

Contribution

We approached Dr. Yehezkeli to assist us with finding an assay to measure produced decursin. We were offered to use NMR which will be faster than HPLC. We were also given input about how should we extract our product from the bacterial culture.

Implementation

Eventually, after discussing the matter with several people from the Yehezeli lab, we decided not to use NMR mainly because using it require larger concentrations than we anticipate, it's noisier in comparison to HPLC and we were able to have a working HPLC program that produces distinct peaks.

Contribution

Our initial concern was that the sample was in a water solution, which isn't HPLC-graded, which results in measurement noise. That's why we used a lyophilizer to freeze-dry the water in the sample, but after running the HPLC, no peaks were obtained.

Dr. Yousef explained to us that freeze-drying a solution may also dry the metabolites in the solution if they have dissolved. Also, the salts in the water won't evaporate and their concentration will increase. He suggested we run the sample and subtract the baseline of water alone. Also, he suggested to try and extract the metabolites using chloroform.

Implementation

Dr. Yousef's help advanced our project and especially the Proof of Concept team in developing appropriate protocols suitable for our metabolites. (see- Protocols page )

With his insight, we were able to overcome the setbacks in our project and came to the conclusion that a different analytical tool is needed (see Measurement page).

As our project progressed, we found out that the detection of our manufactured metabolite, is usually carried out by high-performance liquid chromatography (HPLC). This was not feasible for us due to the high cost and low throughput of the machinery. As we searched for an alternative solutions, we discovered this was a reoccurring problem for many iGEM teams from many tracks: iGEM 2021 HiZJU-China, iGEM 2021 THUSLS China, iGEM HKUST 2021, iGEM ULaval 2018, iGEM EPFL 2020, iGEM Delft 2021 and iGEM NTHU_Taiwan 2016

Implementation

We decided to address this issue and provide an alternative method using the tools available to us in the synthetic biology framework. This resulted in the development of a whole new aspect in our project: the OraCell measurement assay. We propose a new application, which provides a way to quantify decursin. This was achieved by engineering a pathway that exists in mammalian cells. What started as a measurement assay for decursin, could be used by other teams working on the same pathway, or even to mass screen potential new drugs.

Contribution

An integral part of our project was the development of a stable cell line with the Luciferase reporter gene for detecting and quantifying decursin. The development of this assay included transfection of a plasmid which is normally detected using fluorescent activated cell sorting (FACS). This led us to consult with Dr. Aviv Lutaty. We approached him following the transfection of our mammalian cells with a plasmid that contained our bioluminescent protein, Luciferase. Typically, a transfection of this nature is done with a fluorescent protein which is easily identifiable through FACS. However, we chose the more accurate and sensitive bioluminescent protein as opposed to a fluorescent protein. This left us unsure of how to isolate and detect cells that integrated the plasmid.

Implementation

Dr. Aviv Lutaty was very helpful in laying out a clear plan forward for the detection of Luciferase-expressing cells utilizing single cell sorting, fluorescence measurements, and luminescence measurements.

Contribution

As we are looking forward to our project's future, we thought about how we should present the project and its regulation. To overcome this issue, we consulted Dr. Ben David. She offered that we consider how we want to present our product - our target audience should be healthy people with hair loss, and market it as a cosmetic product. This way it will be easier to get approval - and the next step will be to advertise it as a treatment for CIA (after safety checks).

She suggested starting with the US market (when we want to advertise our product), even though their regulation is more complex it will be easier after we get the approval to go to other markets (such as Israel etc.)

Implementation

That conversation helped us to understand better the regulation market and to think about how we want to change the way we think about our product starting from now.

Contribution

Researching about the natural synthesis of decursin in plants taught us that the anabolic pathway from umbelliferone to decursin contains three reactions. Two of them are enzymatic, while the third is a non-enzymatic esterification reaction. We designed our plasmid system to contain genes encoding for the two enzymatic reactions. This meant that we still needed to find a way to include the third reaction from decursinol to decursin in our process. Galit suggested an optional mechanism that includes esterification reaction with an acyl halide and even offered to assist in us in performing it in the wet lab.

Implementation

The reaction that was offered by Galit was the missing piece in our biomanufacturing process. This reaction completes the pathway and yields the wanted metabolite decursin. We plan on performing it experimentally with intentions to integrate it into our biomanufacturing process. Read more in our future plans page.

Professor Yuval Shoham is a researcher in the faculty of Biotechnology and Food Engineering in the Technion

Contribution

When designing our project, we wanted to focus on every step of the way from producing decursin in bacteria to the investigation if our final product would succeed in penetrating to the necessary depth. A part of this process included the modeling of scaling up our biomanufacturing process and its cost. To find out more, visit our modeling page Professor Shoham assisted us in understanding how biomanufacturing projects are typically scaled up and the concerns and costs associated with this process.

Implementation

The biggest piece of advice we took from our conversation was the type of fermentation to be utilized. We debated heavily between batch and continuous fermentation and after our discussion with Professor Shoham, we realized that monetarily and efficiently, batch fermentation was the preferable choice and this is the type we implemented in our model.