䡩杨⵴桲潵杨灵琠浯汥捵污爠瑥捨湩煵敳⁡湤⁰敲獯湡汩穥搠瑲敡瑭敮琠景爠灲業慲礠扲敡獴 捡湣敲 Sjöström, Martin 2018 Document Version: Publisher's PDF, also known as Version of record Link to publication Citation for published version (APA): Sjöström, M. (2018). High-throughput molecular techniques and personalized treatment for primary breast cancer. Lund: Lund University: Faculty of Medicine. 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LUND UNIVERSITY PO Box 117 221 00 Lund +46 46-222 00 00 High-throughput molecular techniques and personalized treatment for primary breast cancer Martin Sjöström, M.D., was born in 1988 in Linköping in Östergötland. Besides working towards MARTIN SJÖSTRÖM precision medicine in oncology, he enjoys training for DIVISION OF ONCOLOGY AND PATHOLOGY | DIVISION OF SURGERY | LUND UNIVERSITY road cycling races and triathlons, building and fying model aircrafts, being in the nature (especially fy fshing or scuba diving), and travelling. He also holds a bachelor’s degree in Latin, with an essay on Virgil’s use of physiological metaphors for psychological phenomena in the Aeneid (Själens spegel – psykets kroppsliga uttryck i Vergilius Aeneiden). *** Precision medicine will transform healthcare by using individualized treatment strategies and approaches. In oncology, this means that the exact changes in individual tumors will be exploited to assess the risk of recurrence, guide treatment decisions, and ultimately targeted with new treatment strategies. This thesis presents fve studies towards better characterization of primary breast cancer by using high-throughput techniques, and further explores how the characterization can lead to individualized adjuvant treatment after surgery. Department of Clinical Sciences Lund Division of Oncology and Pathology Division of Surgery Lund University, Faculty of Medicine Doctoral Dissertation Series 2018:8 ISBN 978-91-7619-575-8 ISSN 1652-8220 MARTIN SJÖSTRÖM High-throughput molecular techniques and personalized treatment for primary breast cancer 2018:8 9 789176 195758 High-throughput molecular techniques and personalized treatment for primary breast cancer 1 2 High-throughput molecular techniques and personalized treatment for primary breast cancer Martin Sjöström Faculty of Medicine Division of Oncology and Pathology, Division of Surgery Department of Clinical Sciences, Lund DOCTORAL DISSERTATION by due permission of the Faculty of Medicine, Lund University, Sweden. To be publicly defended at the lecture hall in the Radiotherapy building, Klinikgatan 5, Skåne University Hospital, Lund, Sweden. January 26th, 2018, 9.00 am. Faculty opponent Professor Dr. John W.M. Martens, Department of Medical Oncology, Erasmus MC Cancer Institute, Rotterdam, The Netherlands 3 Organization: Document name: LUND UNIVERSITY DOCTORAL DISSERTATION Faculty of Medicine Department of Clinical Sciences Lund Division of Oncology and Pathology Date of issue: Division of Surgery 2018-01-26 Sponsoring organization Author: Martin Sjöström Title and subtitle: High-throughput molecular techniques and personalized treatment for primary breast cancer Abstract: Breast cancer is the most common cancer among women worldwide. The heterogeneity between tumors is the basis for precision medicine. Better characterization of tumors, leading to clinical decision tools, is needed to further individualize therapy, and to avoid over-treatment and under-treatment. The work constituting this thesis is focused on personalized treatment of primary breast cancer using molecular high-throughput technologies, and consists of five studies. In study I, we investigated the role of a new putative estrogen receptor, GPR30, in predicting the response to adjuvant endocrine therapy. We showed that endocrine therapy response may be independent of GPR30 expression. On the other hand, the lack of GPR30 expression in the plasma membrane of the cancer cells identified a long-term low-risk group of patients, suggesting a functional change in GPR30 signaling. In study II, we further investigated the functional role of GPR30. We found that it is constitutively active and appears to be pro-apoptotic in its signaling, as well as prognostic for breast cancer outcome. In study III, we evaluated the response to adjuvant radiotherapy after breast-conserving surgery in different breast cancer subtypes. Overall, the subtypes did not appear to be treatment predictive, but the HER2 amplified/over-expressed subtype had the lowest effect of postoperative radiotherapy. We further showed that a presumed low-risk group of patients, similar to patients currently enrolled in clinical trials of de-escalation, have a very good effect of adjuvant radiotherapy on ipsilateral breast tumor recurrences (IBTR), without systemic adjuvant treatment. In study IV, we used gene expression analysis to find tumors that are responsive, or not responsive, to adjuvant radiotherapy. We created a targeted radiosensitivity gene panel, and Single Sample Predictors that were prognostic for IBTR. Combined, they showed promise in stratifying patients for treatment. The correlation of the classifiers with proliferation and immune response could explain the biology behind the models, and may also explain why ours and other classifiers of radiosensitivity perform differently in subgroups of breast cancer. In study V, we used mass spectrometry-based proteomics to search for protein biomarkers of risk of distant recurrence. We created a method of combing shotgun non-targeted discovery mass spectrometry for candidate discovery, and targeted selected reaction monitoring (SRM) mass spectrometry for candidate validation. The method was applied to a cohort of breast tumors first enriched for N-glycopeptides. The workflow was established and a set of 5 candidate protein biomarkers were discovered, which were further orthogonally validated at the gene expression level. In conclusion, the work in this thesis represents a small step towards better personalized treatment of primary breast cancer. Key words: Breast cancer, treatment, high-throughput, gene expression, mass spectrometry, GPR30, GPER Classification system and/or index terms (if any) Supplementary bibliographical information Language: English ISSN and key title 1652-8220 Lund University, Faculty of Medicine ISBN 978-91-7619-575-8 Doctoral Dissertation Series 2018:8 Recipient’s notes Number of pages Price Security clasification I, the undersigned, being the copyright owner of the abstract of the above-mentioned dissertation, hereby grant to all reference sources permission to publish and disseminate the abstract of the above-mentioned dissertation. Signature Date 2017-12-12 4 High-throughput molecular techniques and personalized treatment for primary breast cancer Martin Sjöström Faculty of Medicine Division of Oncology and Pathology, Division of Surgery Department of Clinical Sciences, Lund Supervisor: Dr. Emma Niméus, M.D., Ph.D. Faculty of Medicine Division of Surgery, Division of Oncology and Pathology Department of Clinical Sciences, Lund Co-supervisor: Professor Mårten Fernö, Ph.D. Faculty of Medicine Division of Oncology and Pathology, Department of Clinical Sciences, Lund 5 Coverphoto by Martin Sjöström, arrangement by Jonas Palm. Heatmap of gene expression results. Quod optimus medicus sit quoque philosophus – The best physician is also a philosopher. Title on work by Galenos, originally in Greek, 2nd century AD. Copyright Martin Sjöström Faculty of Medicine Department of Clinical Sciences Lund Division of Oncology and Pathology Division of Surgery ISBN 978-91-7619-575-8 ISSN 1652-8220 Printed in Sweden by Media-Tryck, Lund University Lund 2017 6 To my grandfather, and all cancer victims Quod optimus medicus sit quoque philosophus 7