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Project
Description
ProjectD描述

"Description begins in the writer's imagination, but should finish in the reader's" - Stephen King
“描述始于作者的想象，但应该结束于读者的想象”——斯蒂芬·金

Enlarge image 放大图片

ANCA-associated Vasculitis
ANCA 相关血管炎

Shortcomings 缺点

Our solution 我们的解决方案

Design of !MPACT 设计 ！MPACT

Background information 背景信息

Project 项目

Inspiration 灵感

Future 前途

ANCA-associated vasculitis
ANCA 相关血管炎

Currently, 3-5% of the world population is affected by autoimmune diseases and this number is rising.1,2 Autoimmune diseases are caused by a breach of immunologic tolerance leading to an immune response against molecules of the own body.3
目前，世界人口的 3-5% 受到自身免疫性疾病的影响，而且这个数字还在上升。1,2 自身免疫性疾病是由免疫耐受的破坏引起的，导致对自身身体分子的免疫反应。3


The iGEM TU Eindhoven 2022 team focuses on a group of rare, but life-threatening autoimmune diseases named antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV).4 This is a collection of severe chronic disorders characterized by granulomatous and neutrophilic tissue inflammation causing necrosis of blood vessels.5,6 The necrosis of vessels leads to insufficient oxygen supply to the organs connected to these vessels. As a consequence, a decrease in tissue functioning or tissue death occurs.7
iGEM TU Eindhoven 2022 团队专注于一组罕见但危及生命的自身免疫性疾病，称为抗中性粒细胞胞浆抗体 （ANCA） 相关血管炎 （AAV）。4 这是一系列严重的慢性疾病，其特征是肉芽肿性和中性粒细胞组织炎症导致血管坏死。5,6 血管坏死导致与这些血管相连的器官供氧不足。结果，发生组织功能下降或组织死亡。7


Three types of AAV are classified, namely granulomatosis with polyangiitis (GPA), microscopic polyangiitis (MPA), and eosinophilic GPA (EGPA). As GPA is the most common type of AAV, our project focuses on the chronic, systemic, and relapsing disease GPA.8 GPA predominantly involves the nose and sinuses, lungs, kidneys, joints, and eyes.5
AAV 分为三种类型，即肉芽肿性多血管炎 （GPA）、显微镜下多血管炎 （MPA） 和嗜酸性粒细胞性 GPA （EGPA）。由于 GPA 是最常见的 AAV 类型，我们的项目侧重于慢性、全身性和复发性疾病 GPA。8 GPA 主要涉及鼻子和鼻窦、肺、肾脏、关节和眼睛。5


AAV is associated with the production of pathogenic antineutrophil cytoplasmic antibodies (ANCAs). The ANCA concentration in the body is correlated with the disease activity. These antibodies target two major antigens: leukocyte proteinase 3 (PR3) and myeloperoxidase (MPO).5 The majority of the GPA cases have ANCA autoantibodies targeting PR3.8 These PR3-ANCA-positive patients are at great risk of getting relapses of the disease.7 That is why our project specifically focuses on GPA cases that are PR3-ANCA positive.
AAV 与致病性抗中性粒细胞胞浆抗体 （ANCA） 的产生有关。体内的 ANCA 浓度与疾病活动度相关。这些抗体靶向两种主要抗原：白细胞蛋白酶 3 （PR3） 和髓过氧化物酶 （MPO）。5 大多数 GPA 病例具有靶向 PR3 的 ANCA 自身抗体。8 这些 PR3-ANCA 阳性患者复发的风险很大。7 这就是为什么我们的项目特别关注 PR3-ANCA 阳性的 GPA 案例。

Shortcomings of current treatments
当前治疗方法的缺点

Figure 1 | Shortcomings of the current treatments for AAV. The current treatments for AAV are often immunosuppressive drugs that have multiple shortcomings.
图 1 |目前 AAV 治疗的缺点。目前 AAV 的治疗方法通常是具有多种缺点的免疫抑制药物。
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The current treatments for AAV, and autoimmune diseases in general, are often immunosuppressive drugs, for example, rituximab, cyclophosphamide, or prednisone that suppress the immune system.9 These current treatments are generally effective but due to the unspecific suppression of the immune system, they are associated with burdensome side effects and patients become more prone to infectious diseases.6,10 The cumulative exposure to glucocorticoids and immunosuppressive drugs of current treatments contributes to organ damage, which can cause multimorbidity, i.e. two or more illnesses at a time.9,11 There is a high demand for new treatments that do not contribute to organ damage since AAV already causes major organ dysfunctions.9 Moreover, current treatments can only be used for a limited time as the side effects are burdensome on the patient. As a result, AAV patients often experience relapses of the disease.12 Relapses do not only decrease the patient’s health irreversibly, but also increase the workload of the hospital staff and healthcare costs. The multimorbidity associated with AAV and its treatment increases healthcare expenditures even further.13
目前对 AAV 和一般自身免疫性疾病的治疗通常是免疫抑制药物，例如抑制免疫系统的利妥昔单抗、环磷酰胺或泼尼松。9 目前的这些治疗方法通常是有效的，但由于免疫系统的非特异性抑制，它们与繁重的副作用有关，患者更容易患传染病。6,10 当前治疗的糖皮质激素和免疫抑制药物的累积暴露会导致器官损伤，这可能导致多病共存，即同时出现两种或多种疾病。9,11 由于 AAV 已经导致主要器官功能障碍，因此对不会导致器官损伤的新疗法的需求很高。9 此外，目前的治疗方法只能在有限的时间内使用，因为副作用对患者来说是沉重的负担。因此，AAV 患者经常会出现疾病复发。12 复发不仅会不可逆转地损害患者的健康状况，还会增加医院工作人员的工作量和医疗保健费用。与 AAV 及其治疗相关的多病共存进一步增加了医疗保健支出。13


In conclusion, there is a high unmet need in the treatment of AAV (Figure 1):
总之，AAV 的治疗存在很高的未满足需求（图 1）：

  • They are burdensome9
    他们是累赘的9
  • They are not effective for all patients9
    它们并非对所有患者都有效9
  • They do not exclude relapses9
    他们不排除复发9
  • They cause high healthcare expenditures and workloads13
    它们会导致高昂的医疗保健支出和工作量13

Our solution: !MPACT 我们的解决方案： ！MPACT

To meet the shortcomings of the current therapies against ANCA-associated vasculitis, we designed !MPACT: a Modular and Personalized Autoimmune Cell Therapy. This cell-based therapy detects the ANCA autoantibodies associated with ANCA-associated vasculitis. In the presence of ANCAs, interleukin-10 (IL-10), a natural anti-inflammatory cytokine, gets produced resulting in a specific suppression of the autoimmune response.
为了解决当前针对 ANCA 相关血管炎的疗法的缺点，我们设计了 ！MPACT：一种模块化和个性化的自身免疫细胞疗法。这种基于细胞的疗法可检测与 ANCA 相关血管炎相关的 ANCA 自身抗体。在 ANCA 存在的情况下，会产生白细胞介素 10 （IL-10），一种天然的抗炎细胞因子，导致自身免疫反应的特异性抑制。


!MPACT is based on the Generalized Extracellular Molecule Sensor (GEMS) platform described in Scheller et al. (2018). This modular synthetic receptor allows the coupling of an extracellular input to an intracellular signaling pathway.14 In our case, binding of the pathogenic ANCA autoantibodies to the receptor leads to receptor dimerization and activation, starting a signaling pathway that results in the production and local release of anti-inflammatory cytokine IL-10 (Figure 2). Local release of IL-10 is suggested to have great potential in treating autoimmune diseases according to both literature and stakeholder interviews (see IL-10 in autoimmune diseases and Human Practices).
!MPACT 基于 Scheller 等人 （2018） 中描述的广义细胞外分子传感器 （GEMS） 平台。这种模块化合成受体允许细胞外输入与细胞内信号通路偶联。14 在我们的案例中，致病性 ANCA 自身抗体与受体的结合导致受体二聚化和激活，启动一个信号通路，导致抗炎细胞因子 IL-10 的产生和局部释放（图 2）。根据文献和利益相关者访谈，表明 IL-10 的局部释放在治疗自身免疫性疾病方面具有巨大潜力（参见自身免疫性疾病和人类实践中的 IL-10）。


Figure 2 | Animation of !MPACT cells. ANCAs activate the receptor upon binding which eventually results in the production of IL-10.
图 2 |动画 ！MPACT 细胞。ANCA 在结合时激活受体，最终导致 IL-10 的产生。

The activity of !MPACT is dependent on the concentration of ANCAs present. Higher ANCA concentrations, i.e. higher disease activity, will lead to more IL-10 production, resulting in stronger suppression of the inflammation. Furthermore, the presence of ANCAs is strictly necessary for IL-10 production. When no ANCAs are present, i.e. no disease activity, the cells are “sleeping” and thus will not release IL-10. The production of IL-10 is only desired when ANCAs are present, as elevated IL-10 levels could lead to side effects in the absence of disease.15 In case of a relapse of the disease, the increase in ANCA concentration again activates the !MPACT cells. This way, the relapse of the disease is prevented before it gets problematic. !MPACT is temporarily active, adjusted to the disease activity, and can intervene early on in relapses.
的活动 ！MPACT 取决于存在的 ANCA 浓度。较高的 ANCA 浓度，即较高的疾病活动度，将导致更多的 IL-10 产生，从而更强地抑制炎症。此外，ANCA 的存在对于 IL-10 的产生是绝对必要的。当不存在 ANCA 时，即没有疾病活动，细胞处于“休眠”状态，因此不会释放 IL-10。只有当存在 ANCA 时才需要产生 IL-10，因为在没有疾病的情况下，IL-10 水平升高可能会导致副作用。15 如果疾病复发，ANCA 浓度的增加再次激活 ！MPACT 细胞。这样，就可以在疾病出现问题之前防止疾病复发。!MPACT 是暂时活跃的，根据疾病活动进行调整，并且可以在复发的早期进行干预。


The implementation of !MPACT is similar to CAR T-cells: harvested immune cells from a patient will be genetically engineered with the synthetic receptor and multiplied, after which the cells will be injected back into the patient's body (see Proposed Implementation). In the body, the engineered cells will function as described above to fight the inflammation.
的实现 ！MPACT 类似于 CAR T 细胞：从患者身上收获的免疫细胞将与合成受体进行基因工程改造并繁殖，然后将细胞注射回患者体内（参见拟议实施）。在体内，工程细胞将如上所述发挥作用以对抗炎症。


!MPACT is innovative and powerful since its therapeutic effect is personalized to the patient, i.e. the amount of IL-10 production adjusts to the disease activity. Consequently, fewer burdensome side effects are expected compared to current treatments. In addition, !MPACT can detect and intervene relapses early on, which is not possible with the current treatments. The prevention of relapses and fewer side effects will reduce organ damage and will thereby reduce both healthcare costs and workload. This is confirmed by the Maastricht University Medical Center (see Human Practices). Therefore, we are convinced that our design will greatly contribute to a better treatment of ANCA-associated vasculitis.
!MPACT 具有创新性和强大性，因为它的治疗效果因患者而异，即 IL-10 的产生量根据疾病活动进行调整。因此，与目前的治疗方法相比，预计沉重的副作用更少。另外！MPACT 可以及早发现并干预复发，这在目前的治疗方法中是不可能的。预防复发和减少副作用将减少器官损伤，从而降低医疗保健成本和工作量。马斯特里赫特大学医学中心证实了这一点（参见人类实践）。因此，我们相信我们的设计将极大地有助于更好地治疗 ANCA 相关血管炎。

Design of !MPACT 设计 ！MPACT

Figure 3 | Mechanism of !MPACT cells. The synthetic receptor gets activated upon binding of ANCAs to the PR3 domains, resulting in dimerization of the affinity domains and a rotation of the receptor subunits. Consequently, the downstream JAK/STAT signaling pathway gets induced, resulting in the production of IL-10.
图 3 |的机制 ！MPACT 细胞。合成受体在 ANCA 与 PR3 结构域结合后被激活，导致亲和结构域的二聚化和受体亚基的旋转。因此，下游 JAK/STAT 信号通路被诱导，导致 IL-10 的产生。

The GEMS platform used in our proof of concept cell therapy consists of a mutated erythropoietin receptor (EpoR) fused to an extracellular domain containing proteinase 3 (PR3), to which ANCAs can bind. In addition, the GEMS platform contains signal transduction domains of interleukin 6 receptor B (IL-6RB) that can activate the JAK/STAT signaling pathway.
我们的概念验证细胞疗法中使用的 GEMS 平台由突变的促红细胞生成素受体 （EpoR） 组成，该受体与含有蛋白酶 3 （PR3） 的细胞外结构域融合，ANCA 可以结合该结构域。此外，GEMS 平台包含白细胞介素 6 受体 B （IL-6RB） 的信号转导结构域，可激活 JAK/STAT 信号通路。


Upon binding of the ANCAs to the PR3 domain, the extracellular affinity domains dimerize, causing a rotation of each receptor subunit around its axis. This results in a changed orientation of the intracellular IL-6RB domains, which in turn induces the JAK/STAT pathway. The JAK/STAT pathway results in downstream formation of STAT3 dimers which induce transcription of an IL-10 gene regulated by a STAT3 promoter. After translation of IL-10, the IL-10 is automatically secreted by the cell due to the signal sequence Igk. The mechanism is shown in Figure 3.
ANCA 与 PR3 结构域结合后，细胞外亲和结构域二聚化，导致每个受体亚基绕其轴旋转。这导致细胞内 IL-6RB 结构域的方向发生变化，进而诱导 JAK/STAT 通路。JAK/STAT 通路导致 STAT3 二聚体的下游形成，该二聚体诱导受 STAT3 启动子调节的 IL-10 基因的转录。IL-10 翻译后，由于信号序列 Igk，IL-10 由细胞自动分泌。该机制如图 3 所示。

Interleukin-10 (IL-10) is a cytokine involved in regulating the immune response. Its primary role is to deactivate or dampen the immune response without damaging surrounding tissue.19 This is done by suppressing antigen-specific immunity and thus allowing for T-cell tolerance.20 Since IL-10 lowers the inflammatory host response, it is co-responsible for preventing inflammation in autoimmune pathologies, hence why IL-10 deficiency or dysregulation is also known to be the cause of several autoimmune diseases. The powerful anti-inflammatory characteristics of IL-10 suggest the therapeutic potential of (artificial) IL-10 administration in autoimmune diseases.15 Also interviews with Utrecht University Medical Center, Novartis, and Catherina Hospital showed that IL-10 could have a promising therapeutic effect in autoimmune diseases due to its anti-inflammatory properties (see Human Practices). Administration of IL-10 is especially promising in AAV where the IL-10 production is decreased in case of the development of a relapse.21 This supports the power of !MPACT to prevent relapses of AAV.
白细胞介素 10 （IL-10） 是一种参与调节免疫反应的细胞因子。它的主要作用是在不损害周围组织的情况下使免疫反应失活或抑制。19 这是通过抑制抗原特异性免疫，从而允许 T 细胞耐受来实现的。20 由于 IL-10 降低炎症宿主反应，因此它共同负责预防自身免疫性疾病的炎症，因此 IL-10 缺乏或失调也被认为是几种自身免疫性疾病的原因。IL-10 强大的抗炎特性表明（人工）IL-10 在自身免疫性疾病中的治疗潜力。15 此外，对乌得勒支大学医学中心、诺华和凯瑟琳娜医院的采访表明，由于其抗炎特性，IL-10 在自身免疫性疾病中可能具有有希望的治疗效果（见人类实践）。IL-10 的给药在 AAV 中特别有希望，因为在复发的情况下，IL-10 的产生会减少。21 这支持 了 ！MPACT 防止 AAV 复发。


Although the potential of IL-10 in a clinical application has been demonstrated, many challenges still remain. A significant drawback, which currently withholds clinicians from using IL-10, is that the biologically active form of such a molecule is an unstable homodimer. As a result, IL-10 has a short half-life and is easily degraded in vivo.22 To use IL-10 as a potential therapeutic agent, it is, therefore, necessary to continually administer small doses to maintain the therapeutic concentration. Furthermore, research has shown that, although small concentrations are safe and overall well tolerated, high doses of IL-10 are associated with systemic side effects like fever, headache, and malaise.23 Most of these side effects however are temporary, mild to moderate, and disappear after the cancelation of the treatment.24 The last difficulty that presents itself is the complexity of the IL-10 mechanism. New research suggests that IL-10 is not solely immune inhibitory, but can also stimulate the immune response in some immune diseases and cancers.24,25 A proposed explanation is that IL-10 also induces pro-inflammatory cytokine production, which may be heightened in presence of other proteins, like disease-specific antibodies.26 This is an unwanted effect when using IL-10 as a therapeutic agent. However, much remains uncertain about this effect of IL-10.27
尽管 IL-10 在临床应用中的潜力已经得到证明，但仍存在许多挑战。目前阻止临床医生使用 IL-10 的一个显着缺点是这种分子的生物活性形式是不稳定的同型二聚体。因此，IL-10 的半衰期短，在体内容易降解。22 因此，要将 IL-10 用作潜在的治疗剂，必须持续施用小剂量以维持治疗浓度。此外，研究表明，尽管低浓度的 IL-10 是安全的并且总体耐受性良好，但高剂量的 IL-10 与发烧、头痛和不适等全身副作用有关。23 然而，这些副作用中的大多数是暂时的，轻度至中度的，并在取消治疗后消失。24 出现的最后一个困难是 IL-10 机制的复杂性。新的研究表明，IL-10 不仅具有免疫抑制作用，还可以刺激某些免疫疾病和癌症的免疫反应。24,25 一种可能的解释是 IL-10 还诱导促炎细胞因子的产生，在存在其他蛋白质（如疾病特异性抗体）的情况下，这种细胞因子可能会增加。26 当使用 IL-10 作为治疗剂时，这是一种不良影响。然而，关于 IL-10 的这种作用仍有很多不确定性。27


Despite these disadvantages, local administration of IL-10 has proven to be effective in multiple studies.28-30 Small IL-10 concentrations are shown to be safe, well-tolerated, and effective in the treatment of several autoimmune diseases. Only too high IL-10 concentrations should be prevented since that is associated with pro-inflammatory and systemic side effects.23 To prevent too high IL-10 concentrations, we suggested an inhibition system that can be implemented in !MPACT to ensure safe IL-10 concentrations (see Proposed Implementation).
尽管存在这些缺点，但 IL-10 的局部给药已在多项研究中被证明是有效的。28-30 小浓度的 IL-10 被证明是安全的、耐受性良好的，并且在治疗多种自身免疫性疾病方面是有效的。只应防止过高的 IL-10 浓度，因为这与促炎和全身副作用有关。23 为了防止 IL-10 浓度过高，我们建议使用一种可以在 ！MPACT 确保安全的 IL-10 浓度（参见拟议实施）。

The Generalized Extracellular Molecule Sensor (GEMS) platform is a novel synthetic receptor with great potential for many applications in synthetic biology and for developing cell-based diagnostics and therapeutics. The GEMS platform senses extracellular molecules by modified erythropoietin receptor (EpoR) dimers that are inert for erythropoietin but are fused to extracellular affinity domains which consist of single-chain variable fragments (scFvs) of antibodies, enabling the receptor to sense soluble molecules.14 These EpoR dimers are also linked to different intracellular domains, which transmit the signal and activate distinct natural signaling pathways (JAK/STAT, MAPK, PLCG, and PI3K/Akt).17 Upon ligand binding, the extracellular affinity domains dimerize, causing a rotation of each receptor subunit around its own axis. This results in a changed orientation of the intracellular domains, which in turn activates the downstream signaling pathways.14
广义细胞外分子传感器 （GEMS） 平台是一种新型合成受体，在合成生物学和开发基于细胞的诊断和治疗方法方面具有巨大潜力。GEMS 平台通过修饰的促红细胞生成素受体 （EpoR） 二聚体感应细胞外分子，这些二聚体对促红细胞生成素呈惰性，但与由抗体的单链可变片段 （scFv） 组成的细胞外亲和结构域融合，使受体能够感应可溶性分子。14 这些 EpoR 二聚体还与不同的胞内结构域相连，这些结构域传递信号并激活不同的自然信号通路（JAK/STAT、MAPK、PLCG 和 PI3K/Akt）。17 配体结合后，细胞外亲和结构域二聚化，导致每个受体亚基绕其自身轴旋转。这导致细胞内结构域的方向发生变化，进而激活下游信号通路。14


The modular structure of the GEMS platform allows combinations of different intra- and extracellular domains, making it a customizable epitope sensor. As a result, GEMS are able to detect a large variety of target molecules.14 Until now, the GEMS platform is constructed for the detection of rapamycin, an azo dye (RR120), caffeine, nicotine, a peptide tag fused to mCherry (SunTag), prostate-specific antigen (PSA), and a de novo designed protein displaying two viral epitopes.18 Thereby, the validity of the GEMS platform is confirmed for signaling molecules in a wide range of molecular weights.14 However, it is not yet validated whether the GEMS platform is able to detect antibodies.
GEMS 平台的模块化结构允许不同的细胞内和细胞外结构域的组合，使其成为可定制的表位传感器。因此，GEMS 能够检测多种靶分子。14 到目前为止，GEMS 平台用于检测雷帕霉素、偶氮染料 （RR120）、咖啡因、尼古丁、与 mCherry 融合的肽标签 （SunTag）、前列腺特异性抗原 （PSA） 和显示两个病毒表位的从头设计蛋白。18 因此，证实了 GEMS 平台对宽分子量范围内信号分子的有效性。14然而，尚未验证 GEMS 平台是否能够检测抗体。


The activation of the receptor is usually quantified by the reporter protein SEAP (human placental secreted alkaline phosphatase), making it suitable for detection purposes. Exchanging the reporter protein SEAP for the expression of a therapeutic agent could give rise to multiple other therapeutic applications for the GEMS platform.14

Figure 4 | Schematic overview of the GEMS platform. Both an extracellular affinity domain (a) and an intracellular domain (c) are fused to the EpoR scaffold (b). Different combinations of affinity domains and intracellular domains are possible, making a modular platform. The input molecules cover a wide range of molecular weights, such as Rapamycin, RR120, and PSA. Different signaling pathways (d) can be induced, based on the chosen intracellular domain. All pathways are rewired for transgene expression. At the end of the signaling pathway, a transcription factor will be formed that activates the transcription of a reporter gene (e), or in our case IL-10. The figure is adapted from Scheller et al. (2018)14

Project 项目

During the iGEM project, we designed, built, and tested the proof of concept of !MPACT. Together with stakeholders we validated the problem, designed the therapy, and validated this solution to make the therapy desirable, feasible, safe, and responsible for the world (see Human Practices). The proof of concept is developed and tested in vitro in the lab (see Experiments, Results, and Proof of Concept) Our goal was to show that the designed synthetic receptor could be activated through the binding of antibodies and could subsequently produce IL-10. In addition, we built a kinetic model of the synthetic receptor to support future lab work by simulating the behavior of the system and by predicting sensitive parameters that can be adjusted in the lab to optimize the system and the experimental conditions. This allows the lab work to be used to its full potential (see Model). Furthermore, together with relevant stakeholders in the industry, we created a business plan for !MPACT that allows translation of the project idea directly into a new venture (see Entrepreneurship).


Design goals !MPACT

Click on each circle

Fewer burdensome side effects, by a patient-specific treatment that acts locally in the body

Prevent relapses

Reduce healthcare workloads

General goals of our team

Click on each goal

Project validation

Enlarge public outreach

Professionalization

Collaborations

Sponsoring

Entrepreneurship

As iGEM TU Eindhoven team with many Biomedical Engineering students, we were interested in tackling a global healthcare problem where we could make a real societal impact. First, we started brainstorming about present-day healthcare-related problems and about novel technologies in synthetic biology. We also performed literature research on these topics. Papers suggesting that Long Covid is triggered by an autoimmune response and that the Epstein-Barr virus is the trigger for Multiple Sclerosis sparked our interest. We dived deeper into the topic of autoimmunity. By brainstorming, also together with our PIs, we developed a first concept for our project: designing a synthetic receptor as a treatment for an autoimmune disease. We reached out to several stakeholders, including University Medical Centers, hospitals, and experts in the field of autoimmune diseases, to brainstorm with them about our project idea and to validate the need for new treatments for several autoimmune diseases. All stakeholder interviews can be found on the Human Practices page. Based on the obtained insights from our stakeholders, literature research, and discussions within our team we specified our design. As the involvement of stakeholders in our project was a high priority in our project, we kept validating our project with them throughout the entire journey. Consequently, we had to make several pivots in our project design, but this resulted in a better validated and more desirable design of our project that can eventually make a stronger societal impact.

Future outlook 未来展望

The proof of concept of our modular and personalized autoimmune cell therapy serves as a foundation for a new cell therapy against ANCA-associated vasculitis and for research in synthetic biology. Considering our promising results (see Results), future research should optimize the design of the cell therapy, especially by focusing on enhancing the binding and activation of the GEMS platform by autoantibodies (see Experimental Outlook on the Results page). It is suggested to use a large linker length library to optimize the binding of antibodies and activation of the receptor. Besides, the design could be optimized in order to make the therapy act more locally in the body and the inhibition system for the receptor we envisioned could be implemented to prevent too high IL-10 concentrations (see Proposed Implementation). Next to optimizing the proof of concept, the technology should be patented to protect the IP for future business opportunities (see Entrepreneurship).


The proposed implementation of !MPACT is shown in Figure 5: immune cells will be harvested from the patient, will be genetically engineered with the GEMS platform, and multiplied, after which the cells will be injected back into the patient's body. Before !MPACT can be implemented in the real world, the current proof of concept could be optimized and the treatment should be developed. The development process of the therapy includes performing animal and clinical trials, large-scale production, market authorization, and ensuring reimbursement of the therapy in the basic package of the health insurance (see Proposed Implementation).


Figure 5 | Proposed implementation of !MPACT. First, immune cells are harvested from the AAV patient. These cells are genetically engineered with the synthetic receptor and eventually multiplied. Subsequently, the engineered cells are administered to the patient where the cells will be activated by the ANCA autoantibodies.

!MPACT is currently designed for ANCA-associated vasculitis, however, the modularity of the innovative platform technology offers great potential for this treatment to be applicable to multiple autoimmune diseases. With 400 million people suffering from autoimmune diseases, our technology can eventually help many people and it, therefore, contributes to a healthier future and a reduction in healthcare workloads.

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Fewer burdensome side effects, by a patient-specific treatment that acts locally in the body

As the activation of the synthetic receptor depends on the autoantibody concentration, the therapeutic action will be adjusted to the disease activity. Besides, the goal is that the therapeutic agent IL-10 will be produced locally at the site of inflammation. Consequently, it is expected that fewer side effects will occur compared to the current non-specific immunosuppressive treatments. On the Proposed Implementation page, suggestions are made to achieve the local action of !MPACT.

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Prevent relapses

Ideally, memory cells of the engineered therapeutic cells will be formed to increase the persistence of the therapy in the body. These memory cells can remain in a resting state for years in the human body.16 The memory cells can be activated when a relapse of the disease occurs. This way, the relapse of the disease is prevented before it gets problematic. More information about the memory cells and the implementation of !MPACT can be found on the Proposed Implementation wiki page.

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Reduce healthcare workloads

Since !MPACT will prevent relapses of the disease, it will prevent further health damage and multimorbidity. Consequently, less hospitalization is needed, resulting in lower healthcare expenditures and workloads.

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Project validation

In order to be able to make a societal impact, it is important to validate the problem and the designed solution with stakeholders. Our goal was therefore to involve many relevant stakeholders, including patients, patient foundations, University Medical Centers, hospitals, and pharmaceutical companies throughout the entire project. All input from stakeholders and how we implemented their feedback in our project can be found on the Human Practices wiki page.

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Enlarge public outreach

Since synthetic biology is not yet very well-known at our university and beyond, our goal was to create more awareness for iGEM and synthetic biology in general. Therefore, we focused on public outreach both within our university and outside. This goal is realized by presenting iGEM and the field of synthetic biology in several presentations, social media posts, and publications (see Communication). Besides, we engaged with many stakeholders as well as the general public in our project (see Human Practices, Education, and Partnership). Furthermore, we organized three educational activities about synthetic biology: lessons at elementary schools, a grand Challenge Day for more than 100 High School students, and a workshop and lecture for students at our university (see Education).

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Professionalization

The iGEM Competition is growing and getting more professional, as well as the student teams at the Eindhoven University of Technology. We aimed to follow that trend and wanted to professionalize in order to be able to address and inspire a larger audience to make synthetic biology more well-known. To achieve this goal, we improved the branding of our team, including the use of high-quality media in our outreach (see Communication). Furthermore, the clear organization within our team allowed us to work efficiently and goal-driven. Moreover, we involved relevant experts in our project that together cover all aspects of our project (see Human Practices).

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Collaborations

Collaborating with others can give new ideas and insights which can benefit the project. Our goal was to collaborate with other iGEM teams, our university, and stakeholders, as well as to stimulate others to collaborate. For example, we did not only organize two iGEM meetups to stimulate collaborations between iGEM teams, but we also stimulated collaborations between our partner companies, such as RiboPro, and the university (see Collaborations and Communication).

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Sponsoring

This year’s iGEM TU Eindhoven team focused a lot on getting new sponsorships in order to have enough financial resources to realize all aspects of our project. Finding companies that sponsor our team increases public awareness of the potential of synthetic biology and it indicates that people have faith in the team and project to be successful. All sponsoring collaborations can be found on the page Sponsors.

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Entrepreneurship 创业


To make a real societal impact, our goal is to create a startup out of !MPACT after the iGEM competition. This way, the innovation will be further developed and !MPACT may get on the market eventually. We can thereby help patients suffering from AAV as well as reduce healthcare expenditures and workloads. Therefore, we developed a valid business plan for !MPACT (see Entrepreneurship).

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