1 / 17

R. Graham Cooks, Cedric D’hue, Allison Dill, Demian Ifa,

Hospital Engagement Surgery . R. Graham Cooks, Cedric D’hue, Allison Dill, Demian Ifa, Livia Eberlin, Christina Ferreira, Alan K. Jarmusch Kevin Kerian, Valentina Pirro Department of Chemistry and Center for Analytical Instrumentation Development,

rasha
Download Presentation

R. Graham Cooks, Cedric D’hue, Allison Dill, Demian Ifa,

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Hospital Engagement Surgery R. Graham Cooks, Cedric D’hue, Allison Dill, Demian Ifa, Livia Eberlin, Christina Ferreira, Alan K. Jarmusch Kevin Kerian, Valentina Pirro Department of Chemistry and Center for Analytical Instrumentation Development, Bindley Biosciences Center, Discovery Park, Purdue University Hospital Collaborators: Nathalie Agar, Brigham & Women’s Hospital Timothy Masterson, Michael Koch, Liang Cheng, IUPUI

  2. “Cat on a pillow” DESI Imaging 7 mm m/z 443 Resolution ~ 50 µm

  3. Tissue Analysis : Metabolite Profiling a) One Pixel b) Line Scans c) Tissue Imaging m/z 788 Image construction Signal ratios from MS m/z 448 Tumor margin Justin M. Wiseman, Satu M. Puolitaival, Zoltán Takáts, R. Graham Cooks, Richard M. Caprioli, Angew. Chem. 2005

  4. Brain Tumor Diagnosis by Mass Spectrometry Endogenous lipid profiles Differentiation of tumor subtype, grade, and tumor cell concentration L.S. Eberlin, et al.Proc. Natl. Acad. Sci. 110, 1611 (2013)

  5. Human Brain Cancers • Over 125 types of brain tumors – complex to diagnose • Mean survival 12 – 15 months to some years • New cases (US): 22,910/year, deaths: 13,700/year • Neurosurgery is the primary treatment • Critical decisions made during neurosurgery • Need: Improved intraoperative diagnosis of tumors & margins One Answer: Direct tissue analysis! – Molecular histology using Mass Spec

  6. Human Brain Gliomas – Subtypes Same W.H.O grade (III), different subtypes Gemistocytic L.S. Eberlin, I. Norton, et al Cancer Research, 2012, 72, 645

  7. Human Brain Gliomas – Classification models • A model creation dataset for 3 classifiers was constructed (type, grade, %) using training set • Independent Validation Set Class imaging Results (color coded) L.S. Eberlin, I. Norton, et al Cancer Research, 2012, 72, 645

  8. Prostate Disease State Touch Spray MS ONE PIXEL m/z 700 - 1000 3 years of data; dozens of patients Kevin Kerian, Alan Jarmusch, Valentina Pirro -------Purdue Timothy Masterson, Michael Koch, Liang Cheng--- IUPUI

  9. 788.4 100 885.5 786.4 861.6 812.4 835.3 723.4 816.5 810.3 747.4 Relative Abundance 0 788.4 100 885.5 812.4 834.4 723.5 786.5 810.4 861.5 747.6 0 720 740 760 780 800 820 840 860 880 m/z IUSM – DESI - Prostate PI(38:4) PI(36:4) PS(36:1) 100 788.5 861.5 788.5 PI(34:5) Relative Abundance 0 0.25 0.30 0.35 0.40 0.45 0.50 0.55 0.60 861.5 Time (min) Cancer Normal H&E

  10. MS Pathology: Prostate Alan Jarmusch, Kevin Kerian, Valentina Pirro 12 patients : 100 samples +6 Lack of Analytical (Molecular) Diagnosis Methods Problem Diagnosis: Tumor or Normal Goal DESI-MS Imaging Tool Touch Spray TS-MS DESI Tool (AVG) CV prediction rate = 95% (AVG) CV prediction rate = 96%

  11. Kevin Kerian, Valentina Pirro MS Pathology: Kidney 1 patient Lack of Analytical (Molecular) Diagnosis Methods Problem Diagnosis: Tumor or Normal Goal TS spectrum (avg. 3 scans) TS chronogram DESI-MS Imaging * many peaks – assumed to be adducts Tool 6 seconds *actual photo TS Case 1 TS-MS Tool

  12. Kevin Kerian, Valentina Pirro Alan Jarmusch MS Pathology: Kidney TS Case 1 – directly from surgically resected tissue

  13. DESI-MS Signatures in Bovine Early Embryonic Development Positive ion mode (silver adducts) Negative ion mode Christina Ferreira, PLoS One. 2013 20;8(9):e74981 Immature oocytes Eggs matured in vitro Fertilization Blastocysts developing In vitro Blastocysts Developing In vivo

  14. Ambient Ionization for Early Detection of Parkinson’s Disease in Cerebrospinal fluid • Method for MS/MS for monitoring 33 selected metabolites in 0.47 min of analysis PAPER SPRAY Paper spray quantification using stable isotope internal standards Karen Cesafsky and Ryan Espy Ryan Espy • Reactive paper spray for more sensitivity Derivatization of the primary amine group on amino acids by acetone forming a Schiff base *in ng/mL or ppb; detection in water solutions (no salts present); ** CSF concentration in (in ppb) based on literature reports.

  15. Reactive extraction spray. 18 amino acids can be monitored by MS/MS at CSF concentrations Some amino acids present neutral loss of 46 after acetone reaction and can be detected by NL scanning NL: 8.81E4 129.15 100 blank_acetone_extraction AV: 15 T: + p ESI Full cnl 80 115.01 Ren et al., 2013. Analytical Methods, 5, 6686-6692. 46.000 [80.070-300.000] 60 229.21 157.37 Relative Abundance 143.16 40 178.87 221.02 193.08 117.25 236.14 242.86 201.06 213.04 153.17 183.07 20 245.87 0 172.21 100 NL: 5.43E5 phenyalanine iso/leucine ACSF 1X_acetone_extraction 80 206.03 AV: 25 T: + p ESI Full cnl 46.000 [80.070-300.000] Asparagine valine alanine 60 tyrosine glycine histidine 40 222.84 158.21 184.05 129.22 20 196.09 116.13 0 120 140 160 180 200 220 240 Christina Ferreira and Yue Ren m/z

  16. Parallels between embryo and cancer cell metabolism Immature_1to10_Niemann_TAG_070312 # 61-73 RT: 1.28-1.49 AV: 13 SB: 7 0.03-0.14 NL: 9.57E3 Matured_1to10_Niemann_PL_070312 # 103-138 RT: 3.14-4.20 AV: 36 SB: 14 0.07-0.46 NL: 3.53E2 T: FTMS + p ESI Full ms [600.00-1200.00] T: FTMS - p ESI Full ms [150.00-1000.00] 963.6562 821.5663 100 100 90 90 885.5501 80 80 181.9372 70 70 804.5761 688.1884 60 60 937.6406 786.5291 Relative Abundance Relative Abundance 742.5393 861.5498 50 50 989.6719 625.3921 40 40 738.5221 700.5286 669.4184 30 30 288.8971 713.4444 20 20 266.9151 1015.6879 906.5437 309.0485 394.0259 645.4909 757.4709 801.4969 866.4817 920.7562 10 10 373.8744 1041.7269 479.0038 909.6478 1085.7534 669.1102 1134.5488 584.1326 1173.8081 506.2885 991.5199 0 0 600 650 700 750 800 850 900 950 1000 1050 1100 1150 1200 200 300 400 500 600 700 800 900 1000 m/z m/z • Energy metabolism: increased glycolysis. • Amino acid metabolism: increased glutaminolysis. • Increased fatty acid metabolism: beta-oxidation and de novo synthesis of fatty acids. • Parallels in regulation pathways: pyruvate kinase isoform M2 (PKM2) active in cancer cells and in embryos. • Smith and Sturmey, Biochem. Soc. Trans., 41:664-9 (2013). DESI-MS analysis Embryos are individually placed onto glass slides Mouse or bovine embryos are washed to remove culture media Positive and negative ion mode free fatty acids, cholesterol esters, phospholipids, tryacylglycerols and ubiquinone C.R. Ferreira et al. J Mass Spectrom (2012) C.R. Ferreira et al. Anal. Bioanal. Chem. (2012) Gonzalez-Serrano et al. PLoS One. 2013 Sep 20;8(9):e74981

  17. DESI-MS Ubiquinone as a hallmark in embryo development and cancer stem cells Bovine immature oocyte • ubiquinone ion (of m/z 1140.4, Ag2NO3 adduct) is more abundant in bovine blastocysts compared to immature oocytes. blastocyst produced in vivo High resolution DESI-MS mass spectra of a bovine oocyte (egg) and a blastocyst in the positive ion mode. Abbreviations: TAG – triacylglycerols, Chol ester – cholesterol ester. • ubiquinone adducts appear as major contributors for the differentiation of mouse cells genetically modified to become cancer stem cells (CSCs). Principal component analysis of DESI-MS lipid profiles from control mammary epithelial cells (green circles) and cells genetically modified to express CSCs properties. Valentina Pirro. Collaboration with Alice Chang (Purdue University)

More Related