Team:ZJU-China/Implementation

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Implementation

Product Position

Breast cancer is the major cancerous tumor that harms women's health, with incidence and mortality increasing in lower-income countries. According to the statistics on the Globocon by International Agency for Research on Cancer^[1] in 2020, the incidence rate of breast cancer ranks the first among female tumors worldwide, and the mortality rate ranks the second, second only to lung cancer. In China, breast cancer is one of the most common malignant tumors threatening to female health due to the increasing stress and accelerating pace of life. In the global cancer report released by the international agency for research on cancer (IARC), the cases number of breast cancer in China accounts for 11.19% of the world's, second to United States. In the past few years, both the incidence rate and mortality rate see a makeable increase, which speaks of the significance of comprehensive prevention and control strategies. Since the clinical application of MRI for cancer imaging, there has been a continuous effort in developing tumor-specific contrast agents for accurate cancer detection. To date, there is no targeted contrast agent available for clinical use. The MagHER2some is a disruptive product that fills a market niche and addresses the clinical need of a tumor-specific cancer using MRI. It is now a HER2-positive breast cancer targeted MRI contrast agent, and it would be used in the diagnosis and treatment of many types of human cancers.

Figure 1. Overview of the overall product model.

Industry Background Analysis

Medical Industry Analysis

The main methods of breast cancer screening include physical examination, ultrasound, mammography, magnetic resonance imaging and so on. Mammary examination is divided into breast-self-examination (BSE) and clinical breast examination (CBE). In the United States, the American Cancer Society recommends X-ray screening as the preferred screening method for breast cancer, with women in an average risk of breast cancer receiving regular mammogram screening from age 45, women aged 45 to 54 screening annually, and women aged 55 and over transitioning to every 2 years^[2]. X-ray photography can find clusters of microcalcification and swelling in the breast, which is one of the most effective methods of breast and is widely used in many countries. However, X-ray photography screening also has limitations that may produce false positive and false negative results. Some of the microcalcification biopsies shown by X-rays may be benign lesions^[3], which may not affect survival time without treatment, and the detection of lesions can lead to overtreat. If women are younger and their breasts are denser, X-ray photography may have difficulty distinguishing between microcalculturalization, resulting in a higher false negative rate^[4].

Figure 2. Mammographic molybdenum target detection.

Mammography continues to be the standard imaging modality for screening and work-up of breast cancer^[5]. However, mammography is less sensitive in dense breasts, with reported sensitivity as low as 30%^[6]. Breast magnetic resonance imaging (MRI) is the most sensitive modality in the breast cancer imaging armamentarium to stage breast cancer. Breast MRI outperforms mammography in assessing tumor size^[7]. It also detects additional lesions in 16% to 20% of patients, and diagnoses 3% to 9% of contralateral malignancies^[8]. For high-risk groups, MRI screening has a higher rate of cancer detection than X-ray photography screening, which can detect unexcovered lesions of X-ray photography.

Analysis of MRI Industry

In one research, among 24 cases considered as recurrence by MRI, 23 cases was confirmed by surgical pathology, another case was confirmed by clinical follow-up as misdiagnosis, MRI diagnosis of scar or no recurrence in 28 cases, all of them was confirmed by clinical follow-up or surgery, the sensitivity and specificity of MRI were 100% and 96.5% , as for Ultrasoundgraphy (USG), the data comes as 78.3% and 82.8%. The MRI results is more corresponding with pathology to USG^[9].

Figure 3. Breast MRI.

It is clear that MRI shows a high degree of accuracy in postoperative examination of breast cancer. The Dynamic contrast enhance manifestation differences is the best to distinguish recurrence from scar formation. Combined with the results of our study and related research, the sensitivity and specificity of MRI in breast examinations are superior to that of ultrasound and mammography. MRI can provide valuable information for the identification of postoperative recurrence and scarring. MRI contrast agents have played an imported role for improving the image quality by enhancing the image contrast differences between normal and diseased tissues.

Market and Potential Analysis

According to the population data of the National Bureau of Statistics in 2018 (Figure 4), the total female population in 2018 is 681.87 million, accounting for 48.87% of the total population.

Figure 4. The age structure of China's female population.

From the hospital's point of view, the female population whose age is not covered by the policy is about 336 million. As it is suggested, women over 45 need to do screening annually, and women aged 55 and over transitioning to every 2 years, which is a potential market for MRI with the rapid economic development.

From the point of view of related examination, as people's awareness of health protection continues to increase, the number of women who spontaneously receive breast cancer screening annually is growing rapidly. Based on such demand, the market for targeted MRI in the future will be very broad.

Business Idea

End Users

MRI can help surgeons to obtain adequate surgical margins that ultimately may decrease the local recurrence rate of these tumors.

Our Magnetosome gives the dawn of personalized medicine which clarifies the role of receiving adjuvant chemotherapy and helps doctors to accurately stage breast cancer and determine its local extent^[10].

And we would concentrate radiation delivery at the area closest to the tumor bed, where the risk of recurrence is highest, which will be used after breast conservation surgery^[11].

Implementation Schedule

Figure 5. Implementation Schedule.

Safety

When we had already chosen to develop a project that aims to diagnose HER2 positive breast cancer, we soon realized that MagHER2some is designed for patients in urgent conditions, and we should take strict safety measures to ensure our final product is qualified. To avoid provoking tensions between doctor-patient relationships and make MagHER2some as a accountable product to users and the whole society, we considered and evaluated each safety and ethical issue that might happen in real world.

Safety of Magnetosome BioBricks

The main reason why we chose magnetosome as the most basic BioBricks is that it has been proven to be highly biocompatible^[12]. It has been reported that chemical toxicity of magnetosome from iron ions is negligible^[13][14] due to the insolubility of Fe₃O₄. Therefore, the primary safety issues of magnetosome may be relevant to (a) their nanoscale size, which leads to embolism, blockage, and deposition in the body and (b) impurities, particularly proteins, nucleic acids, and polysaccharides associated with magnetosome extracted from bacterial cells, resulting in immunotoxicity.

As we had a further understanding of those studies related to the biocompatibility of magnetosome, we found out that existing studies have shown the body tissue distribution and host tissue elimination following administration of magnetosome into the vascular system^[15] and in vitro cytotoxicity for mouse fibroblasts^[16].

It has been observed that magnetosome displayed targeted distribution in Sprague-Dawley rat liver except other organs, suggesting that magnetosome may avoid incurring organ congestion or infarction in vivo rely on the lower likelihood of congregation than other nanoparticles.

Purified and sterilized magnetosomes were found to be nontoxic to mouse fibroblasts in vitro. Moreover, another research shows that the injection of 1 mg magnetosome did not increase body temperature of rabbits during the pyrogen test, which showed antigens or pyrogens free with magnetosome administration^[17].

Safety of ZZ Domain BioBricks

ZZ domain is a homologous immunoglobulin-binding domain which belongs to Staphylococcus aureus protein A (SpA). According to our biological engineering, we created a recombinant protein mamC-ZZ which is able to bind to the fragment crystallizable (Fc) region of single chain antibody fragment (scFv) tightly. ZZ domain is a reactive antibody-binding BioBricks, and SpA which ZZ domain belongs to is widely used in clinical application^[18][19], refers to a conclusion that ZZ domain is safe enough to be used as a BioBricks.

Safety of Single Chain Antibody Fragment (scFv) BioBricks

Single chain antibody fragment (scFv) is the most significant important part of our biological engineering design that determines diagnostic efficiency and imaging results of MagHER2some. Accordingly, we chose a certain kind of scFv called modified from a widely use breast cancer medicine Trastuzumab, which has the ability to that binds with high affinity to the extracellular domain of HER2 in a high affinity way. Trastuzumab is a relatively safe and usually well-tolerated drug^[20]. As a monoclonaln antibody, trastuzumab is not modulate by the classic routes of drug elimination or a targeted mode of action^[21]. Thus, problems such as drug-drug interactions and interference with unwanted targets (frequently one of the main causes of drugs 'adverse effects') are unlikely.

Safety of Industry Assurance

Patent and Intellectual Property Protection: We plan to patent MagHER2some after iGEM. Therefore, we have already consulted a law firm for relevant issues.
Strictly Supplement: Considering biosecurity, we only provide the final products to certified medical institutions only. In this way, the possibility of human error, which people misuse or abuse our products, is minimized.
Product Instructions: In terms of a thorough biosafety guarantee, we are going to undertake further clinical trials and upload a informative instructions to illustrate the right execution to every doctor who will use MagHER2some in future.

Advantages

Magnetosome: a Better Choice for MRI Contrast Agent

Compared with artificially synthesized nanosized magnetite, it is the most impressive that magnetosomes have naturally formed phospholipid bilayer membrane. Besides that, magnetosomes have the characteristics of stable single magnetic domain structure, permanent magnetism at room temperature, high chemical purity and uniform particle size and crystal shape^[22]. Taking advantages of these features, we can construct magnetosomes modified with target molecular such as antibodies on its surface to obtain specific contrast agent for different diseases. And compared with chemical synthesis of magnetic nanoparticles, the biological source of the magnetic body with high purity and strong magnetic crystal granularity unique uniform have good biocompatibility and genetic control.

Specificity Demonstration by Recent Research

Surface-modified magnetosomes with a novel peptide (P75) targeting HER2 specifically bind to cancer cells as well as xenograft tumors with surprisingly low accumulation in other organs in vitro and vivo fluorescence imaging results and show obviously negative contrast enhancement in vivo T2-weighted MR imaging^[23], which provides better imaging performance and specificity than existing contrast agents. Different to their design, we chose scFv as our targeting molecular to the HER2-positive tumor cells. The scFv may cause less or even no immune responses. Besides that, based on the diversity of scFv, we can build a platform to design relative antibodies that target different tumors.

Ricks Analysis

Production Ricks and Countermeasures

Competitive Risks and Countermeasures

Technology development and product design will be our key and the inevitable need for long-term development. At present, there are very few products on the market, the market potential is huge.

Financial Risks and Countermeasures

Ethics Risks and Countermeasures

Manage Risks and Countermeasures

Talents are the source of development for our company. The company needs talents in marketing, technology, management, information systems, etc. The core of an enterprise lies in its employees. If the system is not properly managed and management is not timely, the pressure will directly affect corporate culture and indirectly interferes with business operations.

Due to the high price of MRI, the long inspection time, the long product production line and the high cost, in the early days of the company's establishment, there would be few potential customers. We need to continuously improve product performance, win the market with quality, and reduce costs as much as possible to gain market share.

Moreover, we will establish a relatively sound team management system. We will continue to improve team building, from basic research and development to leadership management, to form a complete enterprise system, improve product development, promote innovation and ensure development.

Technical Support

iGEM Community

At the end of August, we were working on the experiments to realize our plan. At the same time, it was also of great importance to hear from others' voices as well as discuss what we like. We participated in the 7th CCiC online meeting, which was focused on the topic, From Lab to Fab, this year. Lectures on transformation of scientific and technology research into fabrication inspired us to explore more outreaches beyond the current project. We were determined to perfect our end product by developing a versatile system from its downstream and upstream after iGEM. In the Q&A part, our biological design to tackle the stability of scFv (a targeting element in our project) got recognition from judges, as well as background investigation. We also had a heated discussion on how to ensure and measure the efficiency of our surface display technology and the possible safety problem we need to address. Constructive insights were freely exchanged in the meeting in order to realize our mutual goals of making a difference in the world with synthetic biology.

Interview with Dr. Fu

We met Dr. Fu in her office in The First Affiliated Hospital, Zhejiang University School of Medicine. Dr. Fu first introduced the issue of breast cancer from the big picture and then in further details, she went on in its standard examination modalities, molecular subtypes and existing therapy. It was after the deep talk with Dr. Fu that we fortunately learned the significance of approaching local medical specialist in breast cancer, who can unveil the local problem clearly and sharply. Accordingly, we shift our orientation from the diagnosis of the disease to the evaluation of post-therapy response and chose to target the HER2 positive breast cancer, the subtype with poor prognosis and malignant rate, in our following works.

Interview with Prof. Chen (radiologist)

We consulted Prof. Chen from The First Affiliated Hospital, Zhejiang University School of Medicine to clear our confusion in MRI. Prof. Chen found it exciting to learn that we focus on the improvement of contrast agent and was interested in the novel design of magnetosome. For one thing, he agreed with our ideas that the magnetosome enclosing iron mineral as active constituent, had lower cellular toxicity and less side effects than traditional agents. He also pointed out the lack of specificity in MRI examination could be the bottleneck of its further application, because much diagnosis today had still been relied on the empirical judgement due to the unspecific imaging. For another, we should consider the size, metabolic model of the magnetosome, for inappropriate size may lead to phagocytosis in liver or fast removal by blood flow. Coincidently, both of us favored the idea of integration of evaluation and treatment by enriching the functions of the mono-function contrast agent.

The dialogue with Prof. Chen contributed to our further consideration into the genetic design to control the structure of magnetosome and its dynamics in modelling process.

Outlook

Contribute to the Real World

If our project wants to contribute to human beings in the real world indeed, MagHer2some must provide more materials on quality control and safety assessment on the existing experimental results until it can be registered and marketed as a medical MRI contrast agent. We will strictly obey the no human experimentation policy, and only discuss the legal, ethical and scientific feasibility of clinical trials that may be carried out in the future.

Clinical Trials Application

At the beginning of our project, we discussed our biological design with Prof. Feng Chen from the imaging department of the First Affiliated Hospital of Zhejiang University. We will conduct experiments on animals with his guidance and help. In vivo experiments are described here, which is conducted scientifically, legally and ethically.

If preclinical studies prove that our product is effective and safe enough, we will go further and apply for clinical trials at the First Affiliated Hospital of Zhejiang University according to the following flow chart:

Framework of Clinical Trials

Magnetic Hyperthermia Therapy

In view of the development trend and concept of the integration of diagnosis and treatment, we hope that the engineering magnetosome as a contrast agent can provide further potential treatment. And then we came up with magnetic hyperthermia.

Magnetic hyperthermia, a form of hyperthermia that is currently undergoing clinical trials. Our magnetosome targeting HER2-positive breast cancer can fully met the requirements of the magnetic hyperthermia medium. In fact, many studies have shown that magnetic nanoparticles as a potential mediator have been proven to be useful in magnetic hyperthermia therapy for a variety of tumors, including breast cancer^[25]-[29]. Therefore, the application of an alternating magnetic field to MNPs could be used in order to induce magnetic hyperthermia and partially or completely destroy small occult lesions limiting the extent of or completely avoiding the need for surgical intervention^[30].

Versatile Targeted Imaging Platform

In the past work, we have successfully developed an engineered contrast agent MagHER2some which specifically targets HER2 positive breast cancer cells^[31]. The core technology we adopted is to display anti-HER2 antibodies on the surface of magnetosome taking advantage of the efficient combination of ZZ domain and Fc region^[32]-[33]. In the same way, we can design diverse scFv modified magnetosomes according to different tumor targets. Looking forward to the future, we aim to develop a versatile targeted imaging platform apply to the clinical diagnosis of more types of tumors (Figure 6).

Figure 6. Specific targets that we can use for different types of tumors.

References

[1]. Bray, F., Ferlay, J., Soerjomataram, I., Siegel, R. L., Torre, L. A., & Jemal, A. (2018). Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: a cancer journal for clinicians, 68(6), 394–424. https://doi.org/10.3322/caac.21492

[2]. Oeffinger, K. C., Fontham, E. T., Etzioni, R., Herzig, A., Michaelson, J. S., Shih, Y. C., Walter, L. C., Church, T. R., Flowers, C. R., LaMonte, S. J., Wolf, A. M., DeSantis, C., Lortet-Tieulent, J., Andrews, K., Manassaram-Baptiste, D., Saslow, D., Smith, R. A., Brawley, O. W., Wender, R., & American Cancer Society (2015). Breast Cancer Screening for Women at Average Risk: 2015 Guideline Update From the American Cancer Society. JAMA, 314(15), 1599–1614. https://doi.org/10.1001/jama.2015.12783

[3]. Johnson, J. M., Dalton, R. R., Wester, S. M., Landercasper, J., & Lambert, P. J. (1999). Histological correlation of microcalcifications in breast biopsy specimens. Archives of surgery (Chicago, Ill. : 1960), 134(7), 712–716. https://doi.org/10.1001/archsurg.134.7.712

[4]. Kolb, T. M., Lichy, J., & Newhouse, J. H. (2002). Comparison of the performance of screening mammography, physical examination, and breast US and evaluation of factors that influence them: an analysis of 27,825 patient evaluations. Radiology, 225(1), 165–175. https://doi.org/10.1148/radiol.2251011667

[5]. Benson, S. R., Blue, J., Judd, K., & Harman, J. E. (2004). Ultrasound is now better than mammography for the detection of invasive breast cancer. American journal of surgery, 188(4), 381–385. https://doi.org/10.1016/j.amjsurg.2004.06.032

[6]. Boyd, N. F., Guo, H., Martin, L. J., Sun, L., Stone, J., Fishell, E., Jong, R. A., Hislop, G., Chiarelli, A., Minkin, S., & Yaffe, M. J. (2007). Mammographic density and the risk and detection of breast cancer. The New England journal of medicine, 356(3), 227–236. https://doi.org/10.1056/NEJMoa062790

[7]. Kuhl, C., Kuhn, W., Braun, M., & Schild, H. (2007). Pre-operative staging of breast cancer with breast MRI: one step forward, two steps back?. Breast (Edinburgh, Scotland), 16 Suppl 2, S34–S44. https://doi.org/10.1016/j.breast.2007.07.014

[8]. Lehman, C. D., Arao, R. F., Sprague, B. L., Lee, J. M., Buist, D. S., Kerlikowske, K., Henderson, L. M., Onega, T., Tosteson, A. N., Rauscher, G. H., & Miglioretti, D. L. (2017). National Performance Benchmarks for Modern Screening Digital Mammography: Update from the Breast Cancer Surveillance Consortium. Radiology, 283(1), 49–58. https://doi.org/10.1148/radiol.2016161174

[9]. Kaifu, W. , Zhanqing, Z. , & Xiesheng, X. U. . (2011). Discussion on the clinical application value of magnetic resonance imaging in fetal abnormality. China Medical Herald.

[10]. Lowery, A. J., Kell, M. R., Glynn, R. W., Kerin, M. J., & Sweeney, K. J. (2012). Locoregional recurrence after breast cancer surgery: a systematic review by receptor phenotype. Breast cancer research and treatment, 133(3), 831–841. https://doi.org/10.1007/s10549-011-1891-6

[11]. Fowble B. (1999). Ipsilateral breast tumor recurrence following breast-conserving surgery for early-stage invasive cancer. Acta oncologica (Stockholm, Sweden), 38 Suppl 13, 9–17. https://doi.org/10.1080/028418699432716

[12]. Sun, J., Li, Y., Liang, X. J., & Wang, P. C. (2011). Bacterial Magnetosome: A Novel Biogenetic Magnetic Targeted Drug Carrier with Potential Multifunctions. Journal of nanomaterials, 2011(2011), 469031–469043. https://doi.org/10.1155/2011/469031

[13]. Häfeli UO, Pauer GJ. In vitro and in vivo toxicity of magnetic microspheres. Journal of Magnetism and Magnetic Materials. 1999;vol. 194(no. 1):76–82.

[14]. Wagner, V., Dullaart, A., Bock, A. K., & Zweck, A. (2006). The emerging nanomedicine landscape. Nature biotechnology, 24(10), 1211–1217. https://doi.org/10.1038/nbt1006-1211

[15]. Sun, J. B., Wang, Z. L., Duan, J. H., Ren, J., Yang, X. D., Dai, S. L., & Li, Y. (2009). Targeted distribution of bacterial magnetosomes isolated from Magnetospirillum gryphiswaldense MSR-1 in healthy Sprague-Dawley rats. Journal of nanoscience and nanotechnology, 9(3), 1881–1885. https://doi.org/10.1166/jnn.2009.410

[16]. Xiang, L., Wei, J., Jianbo, S., Guili, W., Feng, G., & Ying, L. (2007). Purified and sterilized magnetosomes from Magnetospirillum gryphiswaldense MSR-1 were not toxic to mouse fibroblasts in vitro. Letters in applied microbiology, 45(1), 75–81. https://doi.org/10.1111/j.1472-765X.2007.02143.x

[17]. Sun, J., Tang, T., Duan, J., Xu, P. X., Wang, Z., Zhang, Y., Wu, L., & Li, Y. (2010). Biocompatibility of bacterial magnetosomes: acute toxicity, immunotoxicity and cytotoxicity. Nanotoxicology, 4(3), 271–283. https://doi.org/10.3109/17435391003690531

[18]. Navegantes, K. C., de Souza Gomes, R., Pereira, P., Czaikoski, P. G., Azevedo, C., & Monteiro, M. C. (2017). Immune modulation of some autoimmune diseases: the critical role of macrophages and neutrophils in the innate and adaptive immunity. Journal of translational medicine, 15(1), 36. https://doi.org/10.1186/s12967-017-1141-8

[19]. Idowu, O. A., & Heading, K. L. (2018). Type 1 immune-mediated polyarthritis in dogs and lack of a temporal relationship to vaccination. The Journal of small animal practice, 59(3), 183–187. https://doi.org/10.1111/jsap.12774

[20]. Senkus, E., Kyriakides, S., Ohno, S., Penault-Llorca, F., Poortmans, P., Rutgers, E., Zackrisson, S., Cardoso, F., & ESMO Guidelines Committee (2015). Primary breast cancer: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up. Annals of oncology : official journal of the European Society for Medical Oncology, 26 Suppl 5, v8–v30. https://doi.org/10.1093/annonc/mdv298

[21]. European Medicines Agency. Summary of product characteristics: Herceptin. http://www.e ma.europa.eu/docs/en_GB/document_library/EPAR_-_Product_Information/human/000278/WC500074922.pdf. Accessed 9 Feb 2016.

[22]. Uebe, R., & Schüler, D. (2016). Magnetosome biogenesis in magnetotactic bacteria. Nature reviews. Microbiology, 14(10), 621–637. https://doi.org/10.1038/nrmicro.2016.99

[23]. Xiang, Z., Yang, X., Xu, J., Lai, W., Wang, Z., Hu, Z., Tian, J., Geng, L., & Fang, Q. (2017). Tumor detection using magnetosome nanoparticles functionalized with a newly screened EGFR/HER2 targeting peptide. Biomaterials, 115, 53–64. https://doi.org/10.1016/j.biomaterials.2016.11.022

[24]. Lohße, A., Kolinko, I., Raschdorf, O., Uebe, R., Borg, S., Brachmann, A., Plitzko, J. M., Müller, R., Zhang, Y., & Schüler, D. (2016). Overproduction of Magnetosomes by Genomic Amplification of Biosynthesis-Related Gene Clusters in a Magnetotactic Bacterium. Applied and environmental microbiology, 82(10), 3032–3041. https://doi.org/10.1128/AEM.03860-15

[25]. Hu, R., Ma, S., Li, H., Ke, X., Wang, G., Wei, D., & Wang, W. (2011). Effect of magnetic fluid hyperthermia on lung cancer nodules in a murine model. Oncology letters, 2(6), 1161–1164. https://doi.org/10.3892/ol.2011.379

[26]. Zadnik, P. L., Molina, C. A., Sarabia-Estrada, R., Groves, M. L., Wabler, M., Mihalic, J., McCarthy, E. F., Gokaslan, Z. L., Ivkov, R., & Sciubba, D. (2014). Characterization of intratumor magnetic nanoparticle distribution and heating in a rat model of metastatic spine disease. Journal of neurosurgery. Spine, 20(6), 740–750. https://doi.org/10.3171/2014.2.SPINE13142

[27]. Jordan, A., Scholz, R., Maier-Hauff, K., van Landeghem, F. K., Waldoefner, N., Teichgraeber, U., Pinkernelle, J., Bruhn, H., Neumann, F., Thiesen, B., von Deimling, A., & Felix, R. (2006). The effect of thermotherapy using magnetic nanoparticles on rat malignant glioma. Journal of neuro-oncology, 78(1), 7–14. https://doi.org/10.1007/s11060-005-9059-z

[28]. Zhao, Q., Wang, L., Cheng, R., Mao, L., Arnold, R. D., Howerth, E. W., Chen, Z. G., & Platt, S. (2012). Magnetic nanoparticle-based hyperthermia for head & neck cancer in mouse models. Theranostics, 2(1), 113–121. https://doi.org/10.7150/thno.3854

[29]. Kossatz, S., Grandke, J., Couleaud, P., Latorre, A., Aires, A., Crosbie-Staunton, K., Ludwig, R., Dähring, H., Ettelt, V., Lazaro-Carrillo, A., Calero, M., Sader, M., Courty, J., Volkov, Y., Prina-Mello, A., Villanueva, A., Somoza, Á., Cortajarena, A. L., Miranda, R., & Hilger, I. (2015). Efficient treatment of breast cancer xenografts with multifunctionalized iron oxide nanoparticles combining magnetic hyperthermia and anti-cancer drug delivery. Breast cancer research : BCR, 17(1), 66. https://doi.org/10.1186/s13058-015-0576-1

[30]. Ahmed, M., & Douek, M. (2013). The role of magnetic nanoparticles in the localization and treatment of breast cancer. BioMed research international, 2013, 281230. https://doi.org/10.1155/2013/281230

[31] Fiala, O., Pesek, M., Finek, J., Benesova, L., Minarik, M., Bortlicek, Z., & Topolcan, O. (2014). Predictive role of CEA and CYFRA 21-1 in patients with advanced-stage NSCLC treated with erlotinib. Anticancer research, 34(6), 3205–3210.

[32]. Gorantla, B., Asuthkar, S., Rao, J. S., Patel, J., & Gondi, C. S. (2011). Suppression of the uPAR-uPA system retards angiogenesis, invasion, and in vivo tumor development in pancreatic cancer cells. Molecular cancer research : MCR, 9(4), 377–389. https://doi.org/10.1158/1541-7786.MCR-10-0452

[33]. Ragozin, E., Hesin, A., Bazylevich, A., Tuchinsky, H., Bovina, A., Shekhter Zahavi, T., Oron-Herman, M., Kostenich, G., Firer, M. A., Rubinek, T., Wolf, I., Luboshits, G., Sherman, M. Y., & Gellerman, G. (2018). New somatostatin-drug conjugates for effective targeting pancreatic cancer. Bioorganic & medicinal chemistry, 26(13), 3825–3836. https://doi.org/10.1016/j.bmc.2018.06.032

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