War and peace in the tumor microenvironment : tumor-associated cells as facilitators or adversaries during tumor development

The tumor microenvironment, including immune cells, fibroblasts and vasculature, profoundly affects tumor development by initially opposing, but eventually facilitating tumor growth, vascularization and spread. Though corrupted by the growing tumor, such cells remain non-transformed and thus, with proper cues, are possible to direct toward their physiological anti-tumor function. Understanding the mechanisms by which the tumor microenvironment is shaped, before and during tumor growth, has been the principal aim of this thesis. In paper I, we demonstrate that the composition of tumor-associated macrophages (TAMs) can be modulated by selective proliferation of TAM subsets. We identify SEMA3A as a factor potentiating such selective proliferation of anti-tumor TAMs. In paper II we extend the study of SEMA3A’s effect on tumor immunity by showing that it can functionally alter the phenotype of polymorphonuclear myeloid-derived suppressor cells (PMN-MDSC). As a consequence of its effects on TAMs and PMN-MDSCs, the tumor microenvironment is infiltrated by activated cytotoxic lymphocytes which act to obstruct tumor growth. In paper III we show that regulation of mRNA translation shapes the phenotype of TAMs as they become increasingly pro-tumor during tumor growth. We further show that transcripts translationally activated during tumor growth in TAMs were regulated similarly upon M2polarization of macrophages in vitro. Selective inhibition of the MNK2/phospho-eIF4E pathway, which impinges on mRNA translation, functionally altered in vitro M2-polarized macrophages toward a pro-inflammatory phenotype. This suggests that modulation of mRNA translation is a potential target in TAM-based anti-tumor therapies. We further emphasize the importance of mRNA translation in regulating gene expression in the microenvironment in paper IV, where we show changes in its efficiency to drive cancerassociated gene expression alterations in the stroma of patients with chronic obstructive pulmonary disease (COPD). Depending on the lung function, two distinct gene expression programs were discovered. These were enriched for proteins previously identified in fibroblast secretomes that promoted cancer initiation in animal models, highlighting the involvement of non-transformed cells in neoplastic transformation. In paper V, we show that class switch junctions in B cells from patients with BRCA1 mutations display decreased use of non-homologous end joining pathway in favor of the alternative end-joining pathway. This implicates a role for BRCA1 in maintaining genome stability and tumor suppression outside of its recognized role in mediating homologous recombination during cell division. LIST OF SCIENTIFIC PAPERS I. Wallerius M*, Wallmann T*, BARTISH M, Östling J, Mezheyeuski A, Tobin NP, Nygren E, Pangigadde P, Pellegrini P, Squadrito ML, Pontén F, Hartman J, Bergh J, De Milito A, De Palma M, Östman A, Andersson J, Rolny C Guidance Molecule SEMA3A Restricts Tumor Growth by Differentially Regulating the Proliferation of Tumor-Associated Macrophages Cancer Research, 2016, 76(11); 3166-78 II. Wallerius M*, BARTISH M*, Wallman T, Östling J, Joly A-L, Andersson J, Rolny C Semaphorin3A re-educates myeloid derived suppressor cells towards a pro-inflammatory phenotype Manuscript III. BARTISH M, Wallerius M, Wallmann T, Tong D, Liu H, Masvidal L, Joly A-L, van Hoef V, Goncalves C, Seitz C, Bergh J, Del Rincón S, Andersson J, Rolny C**, Larsson O** Translational control of the tumor-associated macrophage phenotype Manuscript IV. Sandri B J*, Masvidal L*, Murie C, BARTISH M, Avdulov S, Higgins LA, Markowski T, Peterson M, Bergh K, Yang P, Rolny C, Limper A H, Griffin T J, Bitterman P B, Wendt C H** and Larsson O ** Distinct cancerpromoting stromal gene expression depending on lung function Under minor revision in American Journal of Respiratory and Critical Care Medicine V. Björkman A, Qvist P, Du L, BARTISH M, Zaravinos A, Georgiou K, Børglum A D, Gatti R A, Törngren T, Pan-Hammarström Q Aberrant recombination and repair during immunoglobulin class switching in BRCA1-deficient human B cells Proc. Natl. Acad. Sci. USA, 2015, 17;112(7):2157-62 ADDITIONAL PUBLICATIONS Not included in this thesis Felgentreff K, Du L, Weinacht KG, Dobbs K, BARTISH M, Giliani S, Schlaeger T, DeVine A, Schambach A, Woodbine LJ, Davies G, Baxi SN, van der Burg M, Bleesing J, Gennery A, Manis J, Pan-Hammarström Q, Notarangelo LD. Differential role of nonhomologous end joining factors in the generation, DNA damage response, and myeloid differentiation of human induced pluripotent stem cells Proc Natl Acad Sci U S A. 2014 Jun 17;111(24):8889-94 *, ** Equal Contribution CONTENTS Foreword . 1 1 Components of the tumor microenvironment . 2 1.1 Tumor-associated macrophages . 3 1.1.1 Macrophage roles in non-pathological physiology . 3 1.1.2 Tumor-Associated Macrophages . 6 1.1.3 Tumor-Associated Macrophage-based therapy strategies . 9 1.2 Myeloid-derived suppressor cells . 11 1.2.1 Ontological origins of MDSCs . 11 1.2.2 The molecular basis of T cell suppression by MDSCs . 12 1.2.3 MDSC-based therapeutic approaches . 13 1.3 Other components of tumor stroma . 14 2 From genotype to phenotype with a focus on mRNA translation . 16 2.1 Discrepancies between mRNA and protein abundance . 17 2.2 Methods to study changes in translational efficiency. 18 2.3 Bioinformatical methods in translatome research . 21 2.4 Cap-dependent mRNA translation initiation . 24 2.5 Regulation of cap-dependant translation initiation . 26 2.5. 1 Regulation of mRNA translation initiation via mTORC1 . 26 2.5. 2 Regulation of mRNA translation initiation by MNK1/2-eIF4E phosphorylation . 28 2.5. 3 Regulation of Ternary Complex formation and the Integrated Stress Response . 29 2.5. 4 Regulation of mRNA translation by RNA-Binding Proteins . 30 2.5. 5 Regulation of mRNA translation by secondary structures in the 3’UTR . 31 2.5. 6 Regulation of mRNA translation by microRNA . 31 3 Thesis findings in context . 32 3.1 Study I . 32 3.2 Study II . 35 3.3 Study III . 37 3.3.1 Translational regulation during TAM progression to a pro-tumor phenotype in vivo . 38 3.3.2 MNK2-mediated effect on M2-phenotype in vitro . 40 3.4 Study IV . 43 3.5 Study V . 46 4 Perspectives and concluding remarks . 50 5 Acknowledgements . 51 6 References . 55 LIST OF COMMONLY USED ABBREVIATIONS