Hydrodynamic Injections: A Case Study

Animal well-being after hydrodynamic injections. In general, the mice tolerated very well the gene transfer procedure via hydrodynamic tail vein injection. All animals recovered from the hydrodynamic injections completely in 5 to 10 min without any signs of distress or problems. The blood creatinine levels did not raise from the pre-hydrodynamic injection baseline values, indicating that the putatively harsh gene transfer method was not harmful to the kidney function, or that one week was long enough to allow kidney function to fully recover from the hydrodynamic stress (Table 1). No statistically significant differences were detected in the creatinine levels of the hydrodynamically injected mice and the non-treated control mice at the analyzed …show more content…
Before the WT diet, all mice showed very similar plasma cholesterol levels (mean total cholesterol 5.3 ± 0.7 mmol/l, SD) and the WT diet induced a substantial increase in the pre-gene transfer cholesterol levels (mean cholesterol 38.6 ± 7.4 mmol/l, SD) (Table 1). Gene transfers with the T2/mLDLR and T2/mVLDLR vectors (in combination with SB100x, or as cis plasmids) to LDLR ko. mice resulted in a trend towards a cholesterol-lowering and stabilizing effect in the treated groups (Fig. 3A). One week after the gene transfers, all the T2/mLDLR and T2/mVLDLR treated mice displayed lowered cholesterol (change in total cholesterol as compared to their individual pre-treatment values (± SEM): -17.4 ± 6.9 %, -6.0 ± 7.2 %, -19.4 ± 3.5 %, and -17.3 ± 5.1 % for the T2/mLDLR, SB100x-T2/mVLDLR(in-one), T2/mVLDLR, and SB100x-T2/mVLDLR(in-one) treatment groups, respectively), with the exception of two individual mice in the SB100x-T2/mLDLR(in-one) treatment group. In contrast, elevated cholesterol levels were observed in all control animals. After the initial declines at one week timepoint, the cholesterol levels of the T2/mLDLR, T2/mVLDLR, and SB100x-T2/mVLDLR(in-one) groups returned to the pre-gene transfer level or were even slightly above it, and continued to rise from thereafter. However, the increase in the cholesterol levels in the treatment groups was moderate in comparison to that seen in the control animals. Some fluctuation was seen in the …show more content…
Prior to the WT diet, all the mice displayed very similar TG levels (mean TG 1.2 ± 0.36 mmol/l, SD). After the gene transfers, the change in TG values did not completely follow the patterns of the cholesterol values (Fig. 3B). However, cholesterol and triglycerides are not metabolized via the same mechanisms. Therefore, the gene therapy approach used herein to increase the LDL-C uptake was not expected to similarly affect the circulating TG levels. No significant differences were found in comparison with the non-transfected controls in the TG levels in a long-term follow-up [at the 16 and 26 weeks’ time points P values were: P = 0.9805 and P = 0.9666 for the T2/mLDLR group; P = 0.9982 and P = 0.9920 for the SB100x-T2/mLDLR(in-one) group; P = 0.9509 and P = 0.9930 for the T2/mVLDLR group; P = 0.9980 and P = 0.4829 for the SB100x-T2/mVLDLR(in-one) group; P = 0.9196 and P = 0.9835 for the T2/lacZ group; P = 0.9996 and P = 0.9961 for the SB100x-T2/lacZ group(in-one),