Q301--Neural Stem Cell Therapy for Spinal Cord cGLP Support Intent to Sole Source

The Veteran s Administration (VA) Office of Research and Development, Regional Procurement Office East in Pittsburgh, PA intends to negotiate on a sole source basis with Virscio, Inc. based in New Haven, CT on behalf of the VA San Diego Healthcare System. The requirement is to provide current Good Laboratory Practice (cGLP)-compliant histopathology services supporting a nonclinical toxicology study evaluating the safety of intraspinal administration of H9 spinal cord neural stem cells (H9-scNSCs) in a cervical hemi-contusion spinal cord injury model in athymic nude rats. The services include tissue processing, paraffin embedding, histologic sectioning, hematoxylin and eosin (H&E) staining, immunohistochemistry, digital slide scanning, pathology evaluation by an American College of Veterinary Pathologists (ACVP)-board-certified veterinary pathologist, quality assurance, and preparation of a final pathology report suitable for inclusion in an FDA Investigational New Drug (IND) application Specific Tasks and Deliverables: 1. Study Support for compliance with GLP documentation requirements SPECIFIC TASK: The Contractor shall provide scientific and administrative support necessary to execute the histopathology portion of the nonclinical safety study. Support shall include study coordination, protocol development and refinement, scheduling personnel and meetings with sponsor, sample tracking throughout each process, communication with the Sponsor, quality oversight of GLP procedures and documentation, and coordination of study activities through completion of the final report. DELIVERABLE: Specimen inventory log including the animal number, treatment group, tissue identification, date received, received by, condition upon receipt, and storage location within 1 week or receipt of tissues. Chain-of-custody documentation noting the transfer of specimens from one location to another within 1 week or receipt of tissue 2. Tissue Trimming received wet SPECIFIC TASK: The Contractor shall receive fixed tissues from a CRO and perform gross trimming of spinal cord, dorsal FboNotice ganglia, peripheral nervous system, and peripheral organ tissues and processed for paraffin embedding (except thoracic and lumbar pieces to be snap frozen and stored in -80C) for subsequent histologic processing. CNS and PNS Tissue Trimming Tissue Location Number of Tissue blocks Section Orientation Analysis Spinal cord Cranial-2 Immediately Cranial to Cranial-1 (~3 mm block) 1 (30% sucrose), transfer to sponsor N/A N/A Spinal cord Cranial-1 Immediately Cranial (~2 mm block) 1 (FFPE) Transverse H&E Spinal cord Contusion site (~1 cm block) 1 (FFPE) Horizontal H&E, STEM121 Spinal cord Caudal-1 Immediately Caudal (~2 mm block) 1 (FFPE) Transverse H&E Spinal cord Caudal-2 Immediately Caudal to Caudal-1 (~3 mm block) 1 (30% sucrose), transfer to sponsor N/A N/A Spinal cord Thoracic 1 (FFPE) Transverse H&E Spinal cord Thoracic 3 (frozen) N/A Snap frozen Spinal cord Lumbar 1 (FFPE) Transverse H&E Spinal cord Lumbar 3 (frozen) N/A Snap frozen DRGs Immediately Cranial 1 (FFPE) Longitudinal N/A DRGs Contusion site 1 (FFPE) Longitudinal N/A DRGs Immediately caudal 1 (FFPE) Longitudinal N/A Medulla Oblongata Whole 1 (FFPE) Transverse H&E *Brachial Plexus Whole 1 (FFPE) Longitudinal N/A Total FFPE blocks = 13 Total FFPE blocks for evaluation = 9 *Brachial Plexus will be collected for the 3-month and 9-month groups. 1-month samples will be stored transferred to molecular CRO for PCR analysis. Peripheral Organ Trimming (Group 1-6, 1 month) Tissue Number of Tissue blocks Analysis 1) Heart 1 H&E 2) Lungs with bronchii 1 H&E 3) Liver 1 H&E 4) Kidney 1 H&E 5) Spleen 1 H&E 6)Testes 1 H&E 7) Ovaries 1 H&E 8) Gross findings if present 1 H&E Total 8 paraffin blocks Thoracic and Lumbar blocks: For all subjects (148 total). From each spinal cord level, 4 pieces of tissue will be trimmed (2-3 mm): Most rostral block will be embedded in paraffin. Other 3 blocks will be snap-frozen and stored. Cranial-1 and caudal-1 blocks: For all subjects (148 total), the cranial and caudal transverse blocks embedded in paraffin will be sectioned in the transverse plane. 12 serial sections (1 slide each) will be sectioned and the remaining block stored. Thoracic and Lumbar blocks (snap-frozen, n=6/rat): Tissue pieces with be shipped on dry ice to molecular CRO for further analysis. All CNS samples tissues will be dehydrated through gradient alcohols, cleared in xylene, and infiltrated with paraffin wax for subsequent embedding. Quantity: 566 tissue specimens DELIVERABLE: Trimmed tissue specimens within 6 weeks of receipt of tissue. Tissue preparation documentation within 6 weeks of receipt of tissue 3. Tissue Trimming received rodent brain SPECIFIC TASK: The Contractor shall perform standardized trimming of rodent brain and medulla specimens according to protocol-defined anatomical landmarks to prepare tissues for paraffin embedding and histologic evaluation. Tissue Location Number of Tissue blocks Section Orientation Analysis Brain Whole 3 (FFPE) Standard H&E Quantity: 150 tissue specimens Trimmed brain specimens within 6 weeks of receipt of tissue Tissue preparation documentation within 6 weeks of receipt of tissue 4. Processing to paraffin blocks SPECIFIC TASK: The Contractor shall process protocol-specified tissues through dehydration, clearing, paraffin infiltration, embedding, and preparation of formalin-fixed paraffin-embedded (FFPE) tissue blocks while maintaining specimen identity and quality control. Quantity: 928 DELIVERABLE: FFPE tissue blocks within 2 weeks of tissue trimming Histology processing records within 2 weeks of tissue trimming Quality control documentation within 2 weeks of tissue trimming 5. Unstained slides SPECIFIC TASK: The Contractor shall section FFPE tissue blocks and prepare unstained microscope slides: CNS/PNS Tissue. Lesion Block: For all subjects (148 total), the lesion block will be sectioned in the horizontal plane. 12 serial sections (1 slide each) will be obtained at each level separated by ~200 µm resulting in at least 6 representative levels through the entire dorso-ventral lesion block. Cranial-1 and caudal-1 blocks: For all subjects (148 total), the cranial and caudal transverse blocks embedded in paraffin will be sectioned in the transverse plane. 12 serial sections (1 slide each) will be sectioned and the remaining block stored. Thoracic and Lumbar blocks: For all subjects (148 total) thoracic and lumbar blocks will be sectioned, 1 slide each. Brain blocks: For all subjects (148 x 3 = 444 total, blocks embedded in paraffin will be sectioned, 1 slide each. Peripheral Tissue For all subjects n=52 from the 1-month timepoint blocks embedded in paraffin will be sectioned, 1 slide each. Quantity 9912 DELIVERABLE: Unstained tissue slides within 8 weeks of paraffin embedding Sectioning logs within 8 weeks of paraffin embedding Slide inventory within 8 weeks of paraffin embedding 6. H&E with slide cutting scanning & pathology evaluation SPECIFIC TASK: Under GLP conditions the contractor shall perform H&E staining of specified tissue sections, and conduct microscopic evaluation by an American College of Veterinary Pathologists (ACVP) board-certified veterinary pathologist. All stains and assays will be performed according to established laboratory methods and microscopically quality checked. Lesion block: 1 slide from each level Cranial-1 and caudal-1 blocks: 1 slide for each block Thoracic and Lumbar blocks: 1 slide for each block Brain blocks: 1 slide for each block Peripheral Tissue: For all subjects n=52 from the 1-month time point 1 slide for each block will be stained for H&E. Quantity: 1166 DELIVERABLE: Histopathology evaluation records withing 12 weeks of paraffin embedding 7. IHC - Manual per marker with scanning and evaluation plus SPECIFIC TASK: The Contractor shall perform manual immunohistochemical staining using protocol-specified antibodies, including the human-specific STEM121 marker, with digital slide scanning and microscopic evaluation by an ACVP board-certified veterinary pathologist. Lesion block: 1 slide from each level will be stained for human-specific STEM121. Quantity: 900 DELIVERABLE: Pathology evaluation records withing 12 weeks of paraffin embedding IHC quality control documentation withing 12 weeks of paraffin embedding 8. Pathology report SPECIFIC TASK: The Contractor shall prepare a comprehensive pathology report summarizing histologic and immunohistochemical findings. The report shall include study methods, microscopic observations, representative images, interpretation of findings, and conclusions regarding the histologic safety of H9-scNSCs. The report shall be suitable for inclusion in an FDA Investigational New Drug (IND) application. DELIVERABLE: Draft pathology report within 8 weeks of slide evaluation Final pathology report within 4 weeks of draft report 9. Study Coordination SPECIFIC TASK: The Contractor shall provide study…