Therapeutic potential of CD22-specific antibody-targeted chemotherapy using inotuzumab ozogamicin (CMC-544) for the treatment of acute lymphoblastic leukemia.

CMC-544 (inotuzumab ozogamicin) is a CD22-specific cytotoxic immunoconjugate of calicheamicin intended for the treatment of B-lymphoid malignancies. This preclinical study investigated antitumor activity of CMC-544 against CD22+ acute lymphoblastic leukemia (ALL). CMC-544 inhibited in vitro growth of ALL cell lines more potently than that of Ramos B-lymphoma cells. When administered to nude mice with established sc xenografts of REH ALL, CMC-544 caused dose-dependent inhibition of xenograft growth producing complete tumor regression and cures in tumor-bearing mice at the highest dose of 160 microg/kg of conjugated calicheamicin. In contrast, a nonbinding control conjugate was 16-fold less effective than CMC-544 in inhibiting growth of REH ALL xenografts. When REH cells were injected intravenously in scid mice and allowed to disseminate systemically, mice developed hind-limb paralysis that was effectively prevented by treatment with CMC-544. Flow cytometric analysis of cells recovered from the bone marrow from mice with disseminated disease verified the presence of engrafted ALL cells. Significantly reduced numbers of ALL cells were recovered from the bone marrow of CMC-544-treated mice than from vehicle-treated mice with disseminated disease. The anti-leukemia activity of CMC-544 demonstrated here further supports clinical evaluation of CMC-544 for the treatment of CD22+ leukemia.

Heat stress increases delivery of a unique sub-population of proteins conveyed by fast axonal transport.

The effect of heat stress on protein synthesis and fast axonal transport was examined in vitro in bullfrog dorsal root ganglion (DRG) and associated spinal/sciatic nerve. Qualitative and quantitative changes of individual 35S-methionine-labelled proteins were determined following DRG labelling and fast transport in respective nerves via two-dimensional gel electrophoresis/autoradiography. Elevation of temperature from 18 degrees C to 33 degrees C for up to 6 hr resulted in a marked increase in synthesis of five individual DRG species of approximately 74,000 daltons that comigrate with heat shock proteins (HSPs). A quantitative comparison of species within this subset revealed two subgroups differentially affected by stress. The three most basic proteins were induced to approximately 1300% of unstressed controls after 6 hr of stress, while the two most acidic species demonstrated an increase to only 300% of controls over the same period. The relative abundance of 25 additional DRG proteins were uneffected by heat stress. Of 70 35S-labelled fast-transported proteins similarly analyzed, 15, comprising 5 families, were consistently transported at greater than 150% of controls following up to 6 hr of heat stress. Over this period all 15 proteins shared a similar profile of abundance relative to non-induced proteins. Transport was elevated to the greatest extent after 2 hr of stress, declined after 3 hr, and tended to rebound at later times. The remaining 55 fast-transported protein spots analyzed were unaffected. An increased delivery of this unique sub-population of 15 fast-transported proteins suggests a possible involvement in early cellular events that mediate heat stress in the nervous system.

Heat stress induces changes in protein synthesis and fast axonal transport in bullfrog sensory neurons.

The effects of heat stress on protein synthesis and fast axonal transport were examined in an in vitro bullfrog primary afferent neuron preparation. The magnitude of effect was determined for individual [35S]methionine-labelled protein species separated via two-dimensional gel electrophoresis. Elevation of temperature of the preparation from 18 degrees C to 33 degrees C caused a transient inhibition of synthesis of non-heat-shock proteins, whereas the synthesis of a 74,000-dalton protein increased to 927% of controls after 4 h. Similar prolonged stress conditions had no effect on the relative abundance of 36 individual, newly synthesized proteins undergoing fast axonal transport. A dramatic exception was represented by a 55,000-dalton glycoprotein whose fast transport was increased to 291% of control. The increase in transport of this protein during a time when synthesis and transport of other non-heat-shock proteins were not enhanced suggests that it may play a unique role in the early cellular events that mediate survival or thermotolerance in the neuron.