Lab of ImmunoEngineering and ImmunoTheranostics VCU School of Pharmacy


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VCU Health. The VCU Health System, which includes the Medical College of Virginia Hospitals, has 1000 inpatient beds and is ranked in the top 100 U.S. hospitals. The VCU Institute for Women’s Health is one of eighteen facilities designated as a National Center for Excellence in Women’s Health by the US DHHS. VCU is closely affiliated with the adjacent Virginia Biotechnology Research Park. VCU also has a Center for Clinical and Translational Research (CCTR), having recently renewed a grant of over 20 million in Clinical and Translational Science from NIH-NCATS (UL1TR002649).

VCU Massey Cancer Center. Massey is among the top 4% of cancer centers in the country and is one of two in the state designated by the NCI to lead and shape America’s cancer research efforts. Founded in 1974, Massey is dedicated to discovering, developing, delivering and teaching effective means to prevent, detect, treat and cure cancer through innovative research, patient care and education. MCC conducts cancer research at every level, including basic science (laboratory), translational, clinical and population sciences research. A major strength for Massey is in facilitating the translation and real-world application of research discoveries into improved treatments and patient care and advances in cancer prevention and control. A key part of the research cycle is developing and conducting clinical trials that test promising scientific breakthroughs, and Massey leads Virginia in offering one of the largest cancer clinical trials menus as well as a statewide clinical trials network that bring Massey’s cutting-edge trials to patients throughout the Commonwealth. Massey is nationally recognized for its work in cancer disparities, studying the socioeconomic and cultural forces causing or contributing to disparities in cancer outcomes with a focus on minorities and the medically indigent. Additionally, Massey provides comprehensive, award-winning cancer care with a full range of medical and support services, the highest-quality treatments, advanced technologies and a compassionate approach. Massey treats patients with all kinds of cancer, including rare and complex cancers. World-class outpatient and inpatient care are delivered at Massey at various sites across the state by multidisciplinary teams of leading specialists who collaborate to fully coordinate and customize each patient’s course of treatment. Massey boasts the state’s largest, most comprehensive bone marrow transplant program and is an internationally recognized pioneer in palliative care. Massey also serves as a vital resource for education, offering academic programs and training for students, cancer researchers and oncology health care professionals as well as health outreach programs for cancer patients, caregivers and the community.

VCU School of Pharmacy. As an integral part of VCU Health, VCU School of Pharmacy ranked #17 nationally by US News (as of 2018). The VCU Department of Pharmaceutics has strong research programs covering nanomedicine and drug delivery, pharmacokinetics and pharmacodynamics, biopharmaceutical analysis, thus making the environment especially rich for drug delivery related research. The close ties and proximity of the Department of Pharmaceutics with other VCU Health components including Massey Cancer Center ensure convenient collaboration between the Zhu Laboratory and Wang Laboratory, Venitz Laboratory, and Poklepovic Laboratory in VCU Health. All VCU laboratories, offices, and core facilities that will participate in this study are either in the same building or within 5-min walk distances from each other (see figure below).  The Zhu Laboratory has full access to various shared state-of-the-art instrumentation in VCU School of Pharmacy located in the Smith Building, including Multi-model microplate reader, multiple flow cytometers, UPLC-mass spectrometers, NMR 400 MHz, cold rooms, and autoclave rooms. The Bioanalytical Laboratory in the Department of Pharmaceutics is equipped with modern instrumentation and procedures necessary for cGLP and FDA guidance compliance. The main equipment in Bioanalytical laboratory includes: LC with triple quadrupole mass spectrometry detection; hybrid triple quadruple/linear ion trap mass spectrometer; GC with mass selective, nitrogen-phosphorous, electron capture and flame ionization detectors; Gamma and Beta scintillation spectroscopy.

Zhu Laboratory: The laboratories (approximately 1,200 sq. ft., 3 rooms) are newly renovated and well-equipped wet lab space, including three main equipment areas for (i) chemistry and formulation development, (ii) biosafety level II tissue culture, and (iii) biological investigation.

Major equipment: The following list of major equipment are available in the Zhu Laboratory, and is fully functional for the proposed work:

  1. 2 chemical fume hoods
  2. 1 BioSearch Expidite DNA/RNA synthesizer
  3. 1 biocompatible Shimazu HPLC for the analysis and the semi-preparative purification of molecular drugs or drug conjugates (with PDA, fluorescence detector of 200 nm - 900 nm)
  4. 1 Labconco freeze dryer
  5. 1 Buchi rotary evaporator
  6. 1 Thermo Scientific Genesys S50 spectrophotometer
  7. 2 magnetic heating pads
  8. 1 scroll pump and 2 oil pumps
  9. 1 water vacuum pump
  10. 1 sonicating water bath
  11. 1 drying oven
  12. 2 balances including a Mettler MTX micro-balance
  13. Multiple benchtop shakers or orbital shakers
  14. 1 BSL II Labconco biological safety cabinet
  15. 2 CO2 cell culture incubators
  16. 1 Barnstead MicroPure di-water purification system
  17. 1 Zeiss AxioVert 200 inverted microscope
  18. 1 Zeiss AxioVert 200 inverted epifluorescence microscope
  19. 1 Thermo Scientific refrigerated centrifuge
  20. 1 water bath
  21. 1 -80 oC freezer
  22. 1 refrigerator
  23. 1 benchtop Legend X3R refrigerated centrifuge
  24. 1 AirClean™ Systems AC600 series vertical laminar flow fume hood (nucleic acid workstation)
  25. 1 dissection microscope
  26. 1 BioRad PCR thermal cycler
  27. 1 BioTek plate washer
  28. 2 horizontal gel electrophoresis systems 
  29. 1 vertical gel electrophoresis
  30. 1 UVP gel imaging system
  31. 1 UV transilluminator
  32. 1 Fujifilm LAS 4000 fluorescence imaging system
  33. 1 Bio-Rad SmartSpec 3000 UV/Vis spectrophotometer
  34. 1 Eppendorf refrigerated centrifuge
  35. 1 pH meter
  36. 1 rotating incubator
  37. 2 benchtop shaker incubators
  38. 2 water baths
  39. 2 refrigerators/freezers
  40. 1 Interscience Scan 300 colony counter

Most Relevant Institutional Resources

VCU maintains state-of-the-art shared facilities and core laboratories to which the PI has full and unrestricted access at minimal charges. These shared facilities, which will be essential to successfully conduct this study, are all within 5-min walk distances from the PI and co-I’s laboratories. These core facilities include the Massey Cancer Center Mouse Model Core (CMMC) Facility, Transgenic/Knock-out Mouse Core, Flow Cytometry Shared Resource Core, Microscopy Shared Resource, Nucleic Acid Core Facility, Molecular Biology Core Facility, and Center for Molecular Imaging (CMI), Tissue and Data Acquisition and Analysis Core (TDAAC), and Molecular Pathology Research Services (MPRS).

Cancer Mouse Model Core: Housed on the first floor of the Massey Building, this core facilitates in vivo translational research by providing assistance with mouse models of cancer and in vivo drug testing. Staff members are experienced in designing and performing experiments using syngeneic, xenograft (including patient-derived xenograft), and genetically engineered mouse models of primary and metastatic cancer. CMMC staff members provide experiment consultation and train researchers in specific animal procedures. The facility is equipped with a cell culture and preparation room that includes a BSL2 cell culture hood, Zeiss Axio Zoom, V16 fluorescence dissection microscope, and BioRad TC20 automatic desktop cell counter; a KOPF 963SD Neurostar Motorized Ultra Precise Small Animal Stereotaxic Instrument for precise delivery of drug or tumor cells; a Xenogen IVIS Spectrum preclinical in vivo imaging system; Vectra Polaris Automated Quantitative Pathology Imaging System, Leica BOND RX automated H&E staining system in the equipment tab, and data management and analysis tools that include StudyLog Desktop software, Living Image 4.5.1 software, ImageJ software, and access to the MCC shared research drive. Using OPAL staining, the Vectra Polaris and Inform software from Akoya Biosciences, the core can perform all the multiplex staining for tumor infiltrating lymphocytes, macrophages, dendritic cells, and myeloid derived suppressor cells.

Transgenic/Knock-out Mouse Core: This core provides a comprehensive suite of services for the creation of genetically modified mice. The facility, located on the 8th floor of the MMRB occupies ~1000 sq ft of space, including a 600 sq ft laboratory, a 150 sq ft tissue culture room, and a 250 sq ft animal holding room. The facility is well-equipped (primary instrumentation includes a Olympus IX-71 inverted injection scope with DP-72 digital camera and stage cooling, Narishige micromanipulators, and a Celtram air microinjector), and offers the production of both transgenic and knock-out or knock-in mice. All stages of transgenic mouse production are offered, including consulting, creation of targeting vectors, ES cell electroporation, screening of ES cell clones, blastocyst injection, and breeding of chimeras. In addition, the core laboratory also has extensive expertise in the mouse line re-derivation by embryo transfer, embryo and sperm cryopreservation, tail DNA preparation and genotyping. The core also maintains an IVIS spectrum imager for live animal imaging, and provides colony management services upon request.

Flow Cytometry Shared Resource Core: It is located in the Molecular Medicine Research Building (MMRB) and provides a comprehensive instrumentation suite and technical services in support of both cell sorting and analysis on a fee-for-service basis. The core currently features seven primary instruments. Cell sorting is conducted using our 5-laser BD FACSAria Fusion sorter as well as our two multi-laser BD Aria II sorters. One of the BD Aria II units is enclosed in a Baker BioProtect Hood, enabling the sorting of human and biohazardous tissues. In addition, the core supports three analyzers, a BD FACSCanto II, a BD LSRFortessa X-20, and an Amnis ImageStreamX Mark II. Sorting capabilities include up to 18 color simultaneous detection (BD FACSAria Fusion) and up to 4 simultaneous sorts per machine. Flow analysis using the X20 allows up to 18 color simultaneous detection. In addition to flow instrumentation, the core maintains a Reichert surface plasmon resonance (SPR) instrument for determination of protein interaction parameters. The resource provides a wide range of services, including instrument training, routine fluorescence analysis, and development of innovative custom analysis and sorting protocols.

Microscopy Share Resource Core: This laboratory is located in an approximately 2500 sq ft facility spread over several rooms on the 9th floor of Sanger Hall, is a fee-for-service core which provides the instrumentation and expertise to facilitate a comprehensive spectrum of imaging methods and techniques. Instrumentation and services include: Electron microscopy (TEM, SEM), Confocal laser scanning microscopy, Spinning disc confocal microscopy, Multi-photon laser scanning microscopy, Structured Illumination microscopy (SIM) (“super-resolution” microscopy), Fluorescence microscopy, Total Internal Reflection Fluorescence microscopy (TIRF), Sample preparation, Live-cell imaging, Immuno-localization (fluorescence, EM), Fluorescence recovery after photobleaching (FRAP), Fluorescence resonance energy transfer (FRET), Stereology, Image analysis.

Tissue and Data Analysis and Acquisition Core Laboratory (TDAAC): Located in the 6,400 sq ft VCU Molecular Diagnostics Facility, this core has as its goal the acquisition of human specimens and associated clinical and pathological findings to support translational research. This is done both through the aegis of the VCU-IRB approved “Tissue Acquisition System to Support Cancer Research” (TASSCR) protocol, which supplies specimens to a biorepository supporting cancer research through acquisition of residual tumor and normal tissue samples along with informed consent from patients with cancer. In addition, TDAAC collects tissue, hematopoietic and serum samples which support specific investigator initiated IRB approved research projects. The specimen acquisition process ensures that the primary purpose of the specimen for patient care is maintained and the quality of the specimen is optimal for biomedical research. These services are achieved through leveraging a network of interdepartmental and informatics relationships within the VCU Health Systems. TDAAC staff in collaboration with Department of Pathology based Molecular Morphology Genomics Laboratory and Laser Capture Microdissection Laboratory provide samples of extracted, quality controlled RNA and DNA from human tissues, frozen sections, and cryopreserved samples of viable hematopoietic neoplasias. Because TDAAC is an outgrowth of a multicenter grant that focused on gene expression microarray studies on multiple cancer phenotypes, some samples have associated gene expression data as a part of their annotation. The VCU Molecular Diagnostics Laboratory is a state-of-the art CLIA-certified laboratory and performs clinical as well as research molecular testing with state-of-the-art equipment, including one bioanalyzer (Agilent 2100), two robotic systems for automated nucleic acid extraction (Roche MagNA Pure LC System), automated DNA sequencers (ABI 310, and 3100), several real-time PCR instrumentation systems (including ABI 7700 and 7500, as well as a Roche LightCycler™ System), a Thermo Spectrophotometer; a Leica CM1850 Cryostat, four Thermo Scientific Revco PLUS Ultra-Low Temperature Freezers, three Thermo Scientific Locator 4 PLUS Cryobiological Storage Vessels, a NanoDrop ND 1000 Spectrophotometer, an Applied Biosystems MagMAX Express nucleic acid extractor, and a complete Affymetrix GeneChip® workstation for DNA microarray technology. All temperature-sensitive equipment is constantly monitored by an electronic system (CheckPoint: 24/7 monitoring system) and all ultra-low freezers have a built-in LN2 backup system that automatically releases LN2 into the freezer in case of temperature failure.

Molecular Pathology Research Services (MPRS): The VCU Molecular Pathology Research Services and Molecular Diagnostics Laboratory is a state-of-the art CLIA’88-certified laboratory and performs clinical as well as research molecular testing with state-of-the-art equipment, including one bioanalyzer (Agilent 2100), two robotic systems for automated nucleic acid extraction (Roche MagNA Pure LC System), automated DNA sequencers (ABI 310, and 3100), several real-time PCR instrumentation systems (including ABI 7700 and 7500, as well as a Roche LightCycler™ System), a Thermo Spectrophotometer; a Leica CM1850 Cryostat, four Thermo Scientific Revco PLUS Ultra-Low Temperature Freezers, three Thermo Scientific Locator 4 PLUS Cryobiological Storage Vessels, a NanoDrop ND 1000 Spectrophotometer, an Applied Biosystems MagMAX Express nucleic acid extractor, and a complete Affymetrix GeneChip® workstation for DNA microarray technology. All temperature-sensitive equipment is constantly monitored by an electronic system (CheckPoint: 24/7 monitoring system) and all ultra-low freezers have a built-in LN2 backup system that automatically releases LN2 into the freezer in case of temperature failure. Additional services include: DNA sequencing of mutational hotspot regions of 50 cancer related genes using next generation sequencing based on AmpliSeq™ technology (Life Technologies, NY).The Oncogenomic Dx One Plus assay detects mutations on selected hotspot regions of 50 cancer genes. Providing support for customized tissue collection, consenting, and sample processing that meet the unique needs of the researcher; providing quality controlled human residual tumor and hematopoietic samples; customized human tissue collection, handling, and informed consent services; and processing of tissue and hematopoietic samples for specialized studies. The VCU TDAAC is supported, in part, by funding from the NIH-NCI Cancer Center Support Grant (P30 CA016059).

Center for Molecular Imaging: The CMI is concerned with multidisciplinary molecular imaging and nanotechnology research using novel approaches in chemistries and multi-modality imaging technologies. The research involves the development of: i) targeted tracers/ probes, and ii) multi-modality imaging strategies to study biological and biochemical pathways in vivo and within the intact environment. The center exploits its imaging science developments and conducts collaborative research with a number of VCU and external investigators, including fostering and facilitating clinical translational imaging research. The center has laboratories for radiochemistry, nano-chemistry and nanotechnology, molecular biology, multi-modalilty animal and tissue imaging. Imaging technologies within the center include preclinical PET/SPECT/CT, optical and MRI/MRS live imaging systems (the facility feature a new Bruker 3T magnet small animal MR machine with a 30 cm bore) as well as micro-autoradiography for ex-vivo imaging. In addition the center is equipped to undertake a complete suite of ancillary methods including optical, ultrasonic, and autoradiographic imaging. Most recently the center has in 2013 acquired a Vevo LAZR photo-acoustic imaging suite.

Nucleic Acids Research Facility and Genomics Core Laboratories: Located in approximately 2000 sq ft on the 5th floor of Sanger Hall, the NARF provides five primary service lines on a fee-for-service basis to VCU researchers. These are: 1) A DNA sequencing core that provides both NextGen and traditional Sanger-based approaches to DNA sequencing and related genomics applications including RNA-Seq/transcriptomics, ChIP-seq, and methylation analysis. NextGen sequencing instrumentation includes a pair of Roche 454 GS-FLX Titaniums, an Illumina HiSeq2500, an Illumina miSeq with cBot workstation, a Life Sciences Proton Torrent, and Covaris nebulizer. Sangernsequencing is supported with an ABI 3730XL sequencer; 2) A DNA microarray core that provides transcriptomics and genotyping/SNP analysis, supported by a suite of instrumentation (fluidics workstation, hybe station, and scanner) supporting Affymetrix DNA arrays, as well as an Illumina Beadstation 500x; 3) A Real-Time PCR core supported by an ABI 7900/HT suitable for Taqman and other popular PCR based assays; 4) A gene synthesis core now specializing in cost-efficient synthesis in FRET probes for Taqman PCR. Recent addition of the 10X Genomics Chromium instruct will allow single cell sequencing analysis. Service arrangements also exist with commercial providers (ACGT inc., BGI Americas, and Eurofins Genomics) for additional needs of NextGen sequencing.

Nanomaterials Characterization Core Laboratory: It provides a comprehensive instrumentation suite for VCU researchers interested in nanomaterials, or surface characterization. The facility features a ThermoFisher ESCAlab 250 X-ray photoelectron spectrometer. The ESCALab is multitechnique platform for studying the surface of materials. The instrument is equipped with a 95 mm field emission gun for Auger spectroscopy and SEM imaging with a 90nm spatial resolution and a UV photoelectron lamp. The sample stage is an automated 5-axis stage for angle resolved XPS with heating and cooling from 77K to 600K. The system also has a High Pressure Gas Cell for introduction of reactive gases for absorption studies. The center also has two scanning electron microscopes and a transmission electron microscope: a Hitachi SU-70 FE-SEM, a Jeol JSM-5610 LV ZEISS libra 120 TEM. The Hitachi SEM is a field emission unit, which allows for a 1nm spatial resolution. The unit also is equipped with a STEM options, a Nabity Lithography System and Genesis EDAX system with low element windows for detection of Be to Pu. The Jeol system is also equipped with an Oxford EDS system and is equipped for High Vacuum SEM as well as Low Pressure SEM functions. The Libra is Zeiss workhorse microscope which features point to point resolution of 0.34 nm. Included in the scope is a in-column Omega filter which allows for energy electron lose spectroscopy (EELS) with an energy resolution of <1.5 eV. The microscopy features a GATAN bottom mount camera for acquisition of digital images. The center also houses a VEECO ICON atomic force microscope. The ICON features VEECO's latest technology providing AFM with less than a 35 pm signal to noise operating in tapping mode. The instrument is capable of operating in multiple modes including lateral force (LFM), magnetic force (MFM), surface potential, scanning capacitance (SCM), Tunneling (TUNA or STM), and conductive (CAFM). The instrument is equipped with a variable temperature stage and a wet cell sample holder.

Biostatistics Shared Resource (BSR): The BSR plays a vital role in supporting research at the cancer center. Faculty members in the BSR are essential collaborators with each of the research programs. The mission of the BSR is to promote excellence in cancer research by providing outstanding biostatistical support and collaboration to Massey members. The defining attribute of this BSR has been to provide biostatistical support to the cancer center researchers beginning in the initial experimental design stage and continuing through all phases, to the presentation of research findings and, ultimately, in the submission of scientific publications. Biostatisticians also participate in the oversight of cancer center protocols in the PRMS and monitoring committees. The emphasis on the collaborative nature of the work of this shared resource has been a major factor in optimizing its effectiveness, resulting in support from peer-reviewed grants. In addition to collaboration at all levels in research projects and grant applications, faculty members of the BSR also contribute through methodological research that applies directly to programmatic research at Massey and in training of investigators through seminars and individual sessions. The BSR also trains postdoctoral scientists and clinical fellows through the clinical research and biostatistics-training track.

Structural Biology Core Laboratory: It facilitates access to a comprehensive suite of instrumentation and computational resources in support of macromolecular structure determination. The shared resource encompasses three related components: X-ray Crystallography, Nuclear Magnetic Resonance, and Molecular Modeling. These facilities are clustered in dedicated space to facilitate access and consultation with the support team. The Structural Biology Shared Resource also collaborates closely with the Biological Macromolecule Shared Resource, which can undertake the production and purification of proteins required for structural analysis. In addition to instrumentation, the core provides training and consultation, both through formal classes, and one-on-one with individual investigators. In 2014 this core added to its capabilities a new suit of instruments for the automation of crystalization conditions. This will include a Gryphon drop setter, Minstrel/Gallery imaging & incubation system for crystallization trials and Alchemist liquid handling system.

Biological Macromolecule Core Laboratory: It provides a range of services to the VCU research community on a fee-for service basis. The focus of the core is the production of biological macromolecules for multiple endpoints. In addition to routine molecular biology support services such as DNA preparations, insert purification, and DNA transformation, value-added services include the production of research grade vector virus particles (adenovirus, adeno-associated virus, retrovirus, and lentivirus), production of targeting vectors for the production of knock-in/knock-out mice (in conjunction with the transgenic mouse core) and somatic cell lines, and other specialized or challenging custom DNA projects. The core also offers an extensive range of protein expression and purification services. Recombinant proteins are available from bacterial, yeast, insect, and mammalian origin. In addition, viral vectors are produced in an access-limited 900 sq ft facility equipped with a Forma double-unit CO2 incubator, a Nikon tissue culture microscope, and a Beckman ultracentrifuge.

The Cancer Research Informatics and Services (CRIS) Core: The CRIS was developed to automate personnel-intensive manual methods for data acquisition, integration and processing through the use of novel informatics systems that support and enhance research activities across VCU Massey Cancer Center. The CRIS Core utilizes four sets of novel cancer informatics software tools and the associated supporting infrastructure to provide necessary services used extensively by Massey researchers. These tools include the Massey Cancer Center Information System (MCCIS), the Automated Cancer Extraction (ACE) application, Clinical Trials Eligibility Database (CTED) and the clinical research data management system (Oncore®), as well as a Universal Datastore, and the recently developed MDalert system. These data and software systems were designed and implemented to meet the informatics and data needs of clinical, basic and population science investigators. The functions of the CRIS Core include data integration, pre-study patient identification for clinical research and custom and standardized report and data set generation used in cohort discovery, grant submission and clinical annotation for tissue and outcomes research. The CRIS Core has focused on the development and support of clinical informatics to a) directly address through automation limitations experienced by investigators and research staff in accessing and processing the data required for evaluating patients for studies, b) integrate, process and store large volumes of disparate clinical data that is not feasible through traditional manual methods, but is essential for performing research; and c) consolidate and link these data in systems that are accessible and useful to investigators. The CRIS Core is closely integrated with the TDAAC and the Clinical Research Shared Resource Core (CRSR).

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