An MRI Evaluation of Patients Who Underwent Treatment with a Cell-Mediated Gene Therapy for Degenerative Knee Arthritis: A Phase IIa Clinical Trial.

Multiple therapies have been developed to slow down the progression of knee osteoarthritis (OA), with the aim of avoiding or delaying TKA. One such potential method is cell-mediated gene therapy, which utilizes allogeneic human chondrocytes modified to express transforming growth factor-ß1. Using magnetic resonance imaging (MRI), we evaluated patients who underwent treatment with this injection in a Phase II study and assessed structural changes in: (1) bone marrow edema lesions, (2) cartilage defect depth and surface area, (3) articular bone surface and osteophytes, and (4) meniscus structure and signal, as well as changes in (5) joint fluid, (6) periarticular inflammation, and (7) synovial inflammation. Twenty-seven patients (6 men and 21 women) who had late-stage OA were randomized 1:1 to receive a 3:1 mixture of nontransduced chondrocytes and genetically engineered chondrocytes, at doses of 6 × 106 cells (group 1) or 1.8 × 107 cells (group 2). MRI was performed at baseline (preinjection), and at 6 and 12 months postinjection. The whole-organ MRI score system was used to assess the aforementioned changes. Treatment was considered to be successful if patients experienced an improvement in or no change in their scores, indicating that the disease had not progressed. All patients in both cohorts individually demonstrated an improvement or no change in one or more of the assessment parameters. At 6 months, the low-dose cohort demonstrated worsening in mean scores in one parameter (bone surface and osteophytes), while the high-dose cohort demonstrated no worsening in mean scores. At 12 months, the low-dose cohort had worsening in the mean score in a subset of one parameter (cartilage signal intensity), and the high-dose cohort demonstrated worsening in mean scores in two parameters (bone surface osteophytes and periarticular inflammation). This is the first study to evaluate MRI changes in patients treated with this injection. These findings provide an impetus for further research on this topic, as well as a starting point for Phase III testing.

A Multicenter, Single-Blind, Phase IIa Clinical Trial to Evaluate the Efficacy and Safety of a Cell-Mediated Gene Therapy in Degenerative Knee Arthritis Patients.

Osteoarthritis leads to articular cartilage wear, and newer therapies are aimed at slowing this degeneration. Growth factors and cytokines influence cartilage formation, and researchers are studying their use on cartilage regeneration in osteoarthritis. One method uses genetically engineered cells to deliver growth factors to damaged cartilage. This technique utilizes transforming growth factor-ß proteins in modified chondrocytes to stimulate cartilage growth via an intra-articular injection. We evaluated the efficacy and outcomes of this injection on patients who had International Cartilage Repair Society grade 4 knee osteoarthritis. We evaluated 27 patients (6 men, 21 women) who had late-stage knee osteoarthritis. Patients were randomized to receive genetically engineered chondrocytes doses of 6×10(6) cells (group 1) or 1.8×10(7) cells (group 2) at a 1:1 ratio. Primary endpoints were subjective and functional evaluations, assessed by the International Knee Documentation Committee (IKDC) score. Secondary endpoints were pain severity and physical function, using the Western Ontario and McMaster osteoarthritis (WOMAC) index and the 100 mm visual analog scale (VAS). Patients were followed at 2, 4, 12, and 24 weeks postinjection. Both groups had significant improvements in outcomes. Scores improved at 12 and 24 weeks from baseline in IKDC (+10 and +14 points in group 1; +11 and +13 points in group 2), WOMAC (-12 and -13 points in group 1; -10 and -12 points in group 2), and VAS (-19 and -24 points in group 1; -20 and -20 in group 2) scores. Additionally, there were no serious adverse events, and no significant difference in adverse event incidence between the groups. Both groups expressed a mean improvement in pain, function, and physical ability following treatment injection. This modality appears to be a promising treatment for cartilage degeneration. However, further larger, multicenter, randomized studies are needed to truly evaluate the efficacy of this novel approach.

Stable integration and functional expression of flounder growth hormone gene in transformed microalga, Chlorella ellipsoidea.

Chlorella is an attractive organism for complex recombinant protein production because of its eukaryotic characteristics and low cost for large-scale culture. Protoplasts of C. ellipsoidea were transformed with a vector containing the flounder growth hormone gene (fGH) under the control of the cauliflower mosaic virus 35S promoter, and the phleomycin resistance Sh ble gene under the control of the Chlamydomonas RBCS2 gene promoter. The presence of introduced DNA was first determined by PCR amplification of both the fGH and Sh ble genes from genomic DNA isolated from transformants and fGH protein expression was detected by immunoblot analysis. Over 400 microg of fGH protein expression per one liter culture containing 1 x 10(8) cells/ml was estimated by ELISA. Stable integration of introduced DNA was confirmed by Southern blot analysis of genomic DNA digested with restriction enzymes. The introduced DNA and fGH expression were detected after seven successive transfers in media devoid of phleomycin, but stably remained in the presence of the antibiotic. Flounder fry fed on the transformed Chlorella revealed a 25% growth increase after 30 days of feeding.