Clinical Tissue Core

Personnel

Overview

The Clinical/Tissue Core serves as an efficient mechanism to provide patient derived "reagents" to investigators associated with the CSIBD. The Core also establishes a reference panel of pedigreed clinical "reagents" for this Center and nationally, as well as serves as a vehicle to promote translational and clinical research. The Core has successfully established a database encompassing IBD patient and an inventory of validated tissues and serum samples, and provides a framework and mechanism linking the clinical resources of the MGH to the needs of researchers with this clinical base.

We follow the conviction that these same assets makes it possible for the CSIBD to fulfill the need broadly recognized by workers in the field for a standard reference panel to validate putative markers of diseases they emerge at the cellular, biochemical, or molecular level in analogy to the role a reference serum panel played in leading to the correct identification of hepatitis C as the agent of non-A, non-B hepatitis. The merits of the approach taken by this Center for a reference bank focusing on tissue, serum and DNA, follows from the two-fold recognition that (1) although genetic factors contribute to IBD, most patients do not have a family history, making it important to have reagents both from members of multiply affected kindreds and patients (and controls) with "sporadic" IBD, and (2) such a resource is most effective in demonstrating an association between a potential disease-related factor and IBD, particularly those which may meet one of the most pressing needs in IBD research, i.e., disease markers.

The Core wants to assure that the basic discoveries of CSIBD investigators (and advancement for elsewhere) are directed toward improvements in patient care. This is achieved by reducing the barriers to entry minimizing the otherwise considerable effort of initiating translational/clinical studies 2) facilitating initiation of collaboration of laboratory based investigators with CSIBD affiliated clinical investigators and an effort to be pro-active in identifying the opportunities for those collaborations.

Core Access

Access to the clinical facilities are easily arranged for CSIBD investigators through Dr. Sands. Dr. Sands also serves as liaison to the GCRC to facilitate its utilization; there is no chargeback by use of the GCRC for use by MGH investigators for research supported by the government. All Center investigators needing tissue samples gain access through Ms. Devaney working under the supervision of Dr. Podolsky. The Core is responsible for procurement of all relevant and suitable samples and it is feasible to accommodate multiple users from surgical specimens. Requests must be submitted in writing to avoid inappropriate tissue handling. Initial requests are limited to twenty-specimens; further request require demonstration of supportive preliminary data derived from the initial series. The guidelines for the amount of tissue and number of samples are provided above in the description of these services. Although conflicts have not proven to be a problem during the current funding interval, if competing demands limit the Core's ability to satisfy all requests, Dr. Lauwers has the responsibility for defining an equitable distribution of materials.

Services

Clinical Database

The Clinical/Tissue Core maintains an active database encompassing patients with IBD seen at the MGH. The database includes salient disease features: symptoms, duration, medications, location of disease (means of documentation), extraintestinal manifestations, endoscopy, surgery, pathology, family history, hospitalizations.

CSIBD investigators with IRB approved projects have access to this data-base through Drs. Sands and Podolsky to identify patients needed for study. An additional responsibility of this Core is the recruitment of patients for specific study needs through notification of all MGH GI Unit practitioners.

Tissue Samples

A primary goal of the Clinical/Tissue Core is to provide tissues and serum for CSIBD investigators and to establish tissue and serum banks. Samples may be dissected or preserved as dictated by the needs of the investigators.

Tissue samples may be provided in several different formats as dictated by the needs of the investigator. These have included harvest in desired buffer, flash frozen in liquid nitrogen, frozen in OCT compound for frozen section and formalin fixation.

CSIBD investigators needing tissue or serum samples contact Dr. Podolsky or Ms. Devaney, Core Technician. She obtains samples as needed from the sources indicated or the stored materials. Clinical validation as needed is provided from the Core data-base through Dr. Sands. CSIBD investigators must have a protocol which has been independently approved by the MGH Subcommittee on Human Studies.

Tissue Reference Bank

In addition to the provision of fresh samples of IBD-related tissues and relevant controls, a main goal of this Core has been to establish a bank of validated tissue and serum samples, maintaining maximum plausible flexibility for later study. All IBD tissues resected at the MGH are available due to the pivotal role of the Co-director.

Guidelines have been developed for utilization of the tissue bank. All investigators must have documentation of Human Studies approval and provide a written request for samples. Investigators may initially receive up to ten individual specimens of a given category and no more than four categories. Further specimens can be requested following review of initial results at the discretion of Dr. Lauwers, the Core Co-Director. Although as noted below, investigators at other institutions may obtain samples from this bank, CSIBD investigators are given priority, with highest priority to recipients of PFS awards.

*Although few surgical specimens of these are available, many have been obtained in smaller tissue quantities through colonoscopic biopsies.

Tissue Sections

The Core provides tissue sections from OCT and paraffin fixed specimens. CSIBD investigators may request up to forty sections (up to five per individual tissue specimen) per year free; additional section will be prepared at a chargeback rate ($10/slide) half the rate charged by the Pathology Department at the MGH. Alternatively, investigators with the capacity may request samples as above to prepare their own sections. Finally, for investigators planning extensive use of prepared sections, training is provided and the investigator may prepare sections using Core equipment.

Serum Bank

The Clinical/Tissue Core ensures the availability of validated serum and blood samples from IBD patients and relevant controls such a resource to CSIBD investigators. Serum samples are obtained from two sources: as noted above, those patients from whom surgical specimens are obtained for the tissue bank and patients as they are enrolled in the clinical database. Input of database information at the time of blood collection ensures detailed corroborative information on these specimens.

The serum bank includes samples from patients with relevant diseases (non-IBD inflammatory disease of the small and large intestine, non-GI inflammatory disease, (e.g., rheumatoid arthritis), neoplastic disease and non-disease controls. This bank also includes serum from IBD patients not undergoing surgery according to the following categories:

• Ulcerative Colitis:
• Active/Non-active
• Universal/Left-sided/Rectum only
• Crohn's Disease
• Active/Non-active
• Small Intestine/Colon/Both/Other
• Treated/Non-treated
• Extraintestinal Manifestations

Center investigators may initially obtain an aliquot (0.5ml) from up to 20 patients for Human Studies approved protocol. Access is limited to Center investigators. Further samples can be obtained based on the results of the analysis of the initial cohort of sera studied.

Immortalized IBD Lymphocytes and DNA

In conjunction with the expanded Core goal of a serum bank of pedigreed samples as outlined above, an additional 10ml of blood has been collected from each of the individuals contributing to the serum sample. Granulocytes and lymphocytes have been collected separately from these samples by Ficoll-hypaque centrifugation. Subsequently, genomic DNA is prepared from granulocytes by routine methods and stored under absolute ethanol. In parallel, lymphocytes are used to establish cell lines by EBV-immortalization using the protocol of Sugden (1). The power of this resource is enhanced by the corresponding serum bank and coordinate clinical database.

Clinical Research Support

The Clinical/Tissue Core provides the means to undertake a complete spectrum of research in patients. Through the clinical resources of the MGH/GI Unit and the newly opened IBD clinical and clinical research facility and General Clinical Research Center of the MGH the following services can be provided:

• Patient Identification and Recruitment (see above)
• Phlebotomy
• Endoscopic evaluation and procurement of tissue specimens
• Inpatient monitoring
• Radionuclide scanning and comprehensive radiologic services
• Bacteriologic Services

The opening of a dedicated Clinical IBD Center at MGH which integrates space for personnel supporting clinical investigation provides a major enhancement of this cores ability to support clinical and translational IBD research.

Biostatistical Analysis

Expertise in study design, data base management and analysis will be available through consultation with Dr. Schoenfeld of the GCRC. Dr. Sands also serves as a resource for this expertise. These interactions will be further supported by access to the computer resources of the MGH/GI Unit and the GCRC which interface with the mainframe computers used by the members of the CSIBD at the MGH. The GCRC provides the CLINFO computerized data base management and analysis system as well as the SAS, SAAM, and PULSAR data analysis programs.

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