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Apprenticeships are perfect if you want to combine theoretical learning with practical, hands-on experience (and a Ambrogio, C. et al. Combined inhibition of DDR1 and Notch signaling is a therapeutic strategy for KRAS-driven lung adenocarcinoma. Nat. Med. 22, 270–277 (2016).

Department of Pathology, School of Medicine and Health Sciences, The George Washington University, Washington, DC, USA

a) Diagram of full-length (FL) DDR1 (top) and tumour curves of either E0771 Ddr1-WT or KO tumour cells carrying various DDR1 expression vectors: empty vector (EV), FL, deletion of the kinase domain (ΔKD), and extracellular domain (ECD) only. All p values were compared to KO + EV group. TM: transmembrane domain. WT: n = 9 tumours, KO+EV: n = 10 tumours, KO+FL: n = 10 tumours, KO+ ΔKD: n = 6 tumours, KO+ECD: n = 5 tumours. ( b) Crystal structure of mouse DDR1 collagen-binding domain, generated by Jmol software ( http://www.jmol.org/). Amino acid residues targeted in the mutational analysis are shown. ( c) Immunoblots of Flag-tagged mouse WT DDR1-ECD and point mutants ectopically expressed in M-Wnt tumour cells, with GAPDH as the loading control. Images are representatives from three independent experiments. ( d) Immunoblots of Flag-tagged mouse WT DDR1-ECD and point mutants ectopically expressed in AT-3 tumour cells, with GAPDH as the loading control. Images are representatives from three independent experiments. ( e–f) Growth curves of M-Wnt (e) and AT-3 (f) Ddr1-KO tumours with ectopically expressed mouse WT DDR1-ECD or collagen-binding point mutants. The numbers in parenthesis indicate outgrowing tumours (larger than 100 mm 3) versus total injected. ( g, h) Immunoblots of full-length DDR1 in cells and soluble ECD in conditioned medium from various mouse (g) and triple-negative human breast cancer cell lines plus ER-positive MCF7 (h). Images are representatives from three independent experiments. ( i) Coomassie staining of recombinant Fc-ECD under non-reducing and reducing conditions. ( j) Rescue of Ddr1-KO E0771 tumour growth in immunocompetent hosts by recombinant Fc-ECD versus PBS vehicle (n = 6 tumours/group). ( k) Diagram of the Transwell assay for CD8 + T cell migration. Primary CD8 + T cells were loaded in the upper chamber that had been pre-seeded with decellularized ECM derived from tumour cells. The lower chamber contained medium with or without CCL21. ( l) CD8 + T cells in vitro migration activity was abrogated by decellularized ECM from AT-3 tumour cells in a DDR1-dependent manner. Value of migrated CD8 + T cell number without ECM and CCL12 is set at “1” (lanes 1 and 2: n = 3; lanes 3 and 4: n = 7), n refers to technical repeats. Values represent mean ± SEM. p value as indicated, two-tailed Student’s t-test for all tests except for tumour volumes, which were done by two-way ANOVA.

a) Representative images of CD8 + T cell staining at E0771 tumour margin and in the tumour core (bottom panels). ( b, c) Representative images (b) and quantification (c) of CD8 + T cell IHC at M-Wnt tumour margin and core (WT: n = 8 tumours, KO: n = 4 tumours). ( d, e) Representative images (d) and quantification (e) of CD8 + T cell IHC at AT-3 tumour margin and core (n = 5 tumours/group). Images in (a),(b), and (d) showing tumour margin at top panel (tumour boarder denoted by red dash lines) and tumour core at bottom panel. Box areas at higher magnification are shown in the upper right inlets. Red arrow heads indicate CD8 + cells. The y-axis in (c) and (e) refers to percent of CD8 + cells over total cells in a given field. Scale bar: 100 µm and 10 µm in inlets. Two-tailed Student’s t-test. ( f–g) Correlation between DDR1 mRNA levels and overall survival of all patients with breast cancer (f) and patients with TNBC (g) in the Kaplan-Meier Plotter database ( https://kmplot.com/analysis/). ( h–j) Scatter plots showing the negative gene expression ( Z-score) correlation between DDR1 mRNA levels and GZMB (h), IFNG (i), and PRF1 (j) in TCGA TNBC tumours (n = 162). The corresponding Spearman’s correlation coefficients and p values are shown. ( k–n) Correlation of DDR1 mRNA levels and anti-tumour immune markers in 37 samples from patients with TNBC (GSE88847). ( o) Scatter plot showing the negative gene expression correlation between DDR1 mRNA levels and signature for accumulation of T cells in tumours using TCGA TNBC tumour data. ( p) Scatter plots showing the negative expression correlation between DDR1 protein expression and cytolytic effector pathway in CPTAC BRCA. ( q) Correlation between percentages of CD8 + immune cells and DDR1 + tumour cells in a TNBC cohort (n = 12). ( r) Correlation between percentages of CD8 + immune cells and DDR1 + tumour cells in a DDR1 high (n = 7) and DDR1 low (n = 5) TNBC samples. ( s) Patient numbers of immune-excluded (n = 4) and non-immune-excluded (n = 6) in DDR1 high and DDR1 low group. Only the 10 patient samples with paired margin and core information were used for the immune exclusion calculation in Extended Data Fig. 3s, two-sided Chi-square test.

Resources

Hanzelmann, S., Castelo, R. & Guinney, J. GSVA: gene set variation analysis for microarray and RNA-seq data. BMC Bioinformatics 14, 7 (2013). Raudvere, U. et al. g:Profiler: a web server for functional enrichment analysis and conversions of gene lists. Nucleic Acids Res. 47, W191–W198 (2019). Baarlink C, Wang H, Grosse R . Nuclear actin network assembly by formins regulates the SRF coactivator MAL. Science 2013; 340: 864–867. Lim KL, Chew KCM, Tan JMM, Wang C, Chung KKK, Zhang Y et al. Parkin mediates nonclassical, proteasomal-independent ubiquitination of synphilin-1: implications for Lewy body formation. J Neurosci 2005; 25: 2002–2009. van de Sluis B, Rothuizen J, Pearson PL, van Oost BA, Wijmenga C . Identification of a new copper metabolism gene by positional cloning in a purebred dog population. Hum Mol Genet 2002; 11: 165–173.

Takahashi Y, Sipp D, Enomoto H . Tissue interactions in neural crest cell development and disease. Science 2013; 341: 860–863.Department of Biochemistry and Molecular Medicine, School of Medicine and Health Sciences, The George Washington University, Washington, DC, USA Edwards, D. N. et al. Selective glutamine metabolism inhibition in tumor cells improves anti-tumor T lymphocyte activity in triple-negative breast cancer. J Clin. Invest. 131, e140100 (2021).

O'Hara A, Simpson J, Morin P, Loveridge CJ, Williams AC, Novo SM et al. p300-mediated acetylation of COMMD1 regulates its stability, and the ubiquitylation and nucleolar translocation of the RelA NF-kappaB subunit. J Cell Sci 2014; 127: 3659–3665. Vogel, W. F., Aszodi, A., Alves, F. & Pawson, T. Discoidin domain receptor 1 tyrosine kinase has an essential role in mammary gland development. Mol. Cell. Biol. 21, 2906–2917 (2001).

You will build your skills over a 14 month period covering:

Newman, A. M. et al. Determining cell type abundance and expression from bulk tissues with digital cytometry. Nat. Biotechnol. 37, 773–782 (2019). Agarwal, G., Mihai, C. & Iscru, D. F. Interaction of discoidin domain receptor 1 with collagen type 1. J. Mol. Biol. 367, 443–455 (2007). Deng ZH, Gomez TS, Osborne DG, Phillips-Krawczak CA, Zhang JS, Billadeau DD . Nuclear FAM21 participates in NF-kappa B-dependent gene regulation in pancreatic cancer cells. J Cell Sci 2015; 128: 373–384.