Thérapie provasculaire anti-cancer à l’aide de microbulles stimulées par ultrasons afin d’augmenter l’efficacité de la radiothérapie

In solid tumors, hypoxia is a recognized mechanism of resistance to radiation therapy. It has recently been shown that ultrasound (US) targeted microbubbles (MBs) cavitation (UTMC) can increase blood perfusion in skeletal muscles by triggering nitric oxide signaling. Interestingly, this effect was amplified with a sodium nitrite co-injection and reduced the spasm observed with high pressure long tone burst ultrasound. Since sodium nitrite has been shown to synergize with radiotherapy, we hypothesized that UTMC with a sodium nitrite co-injection could further radiosensitize solid tumors by increasing blood perfusion and thus reduce tumor hypoxia. We evaluated the ability of UTMC with and without nitrite to increase perfusion in muscle (mouse hindlimbs) and tumors using different pulse lengths and pressure and evaluated the efficacy of this approach as a provascular therapy given directly before radiotherapy treatments. Human prostate xenografts (PC3) were grown bilaterally in immunodeficient mice: one side was treated with UTMC, and the contralateral side was used as a control. Therapeutic transducers (long pulses) or cardiac phased array (short pulses) were used to treat the tissue of interest during the injection of MBs via the tail vein. Nitrite was injected 5 minutes before UTMC when appropriate. UTMC consisted of 60 therapeutic pulses, given at a pulse interval (10-15 seconds) adjusted to allow MB replenishment, as guided by contrast enhanced US imaging (CEUS), and was typically given in 15 minutes. The increase in perfusion in the tumor was quantified by burst replenishment imaging allowing longitudinal quantification of blood perfusion (A×B). Mice were irradiated 10 minutes after UTMC, and tumor growth followed for 25 days. In muscle, the increase in blood perfusion following UTMC treatment with various long pulses was strong. In tumors subjected to long pulses, the increase in perfusion was significant at lower M.I. (125 and 250 kPa) but not at higher M.I. (375, 500 and 750 kPa) when compared to control (0 kPa). When combined with radiotherapy, UTMC with long pulses at 250 kPa did not result in a significant increase in treatment efficacy. UTMC with short pulses (SONOS, 1500 kPa) and nitrite co-injection was then validated in muscle. The effect of treatment was strong and significative. 8 This significant increase in perfusion was also visible in tumors subjected to UTMC (SONOS) + nitrite and lasted for at least 10 minutes but not for nitrite alone. The blunted provascular responses observed for long pulses at higher M.I. without nitrite was reversed with the addition of nitrite. SONOS with and without nitrite caused an increase in perfusion. Histological validation of reperfusion in SONOS + nitrite was also significant when compared to tumors receiving only nitrite alone. Finally, there was an improved growth inhibition for the 8 Gy + nitrite + UTMC group vs. 8 Gy + nitrite alone with the SONOS therapy. This effect was not significant with mice treated by UTMC + nitrite and receiving 0 Gy or 2 Gy. In conclusion, UTMC + nitrite seemed to increase blood flow leading to an increased radiotherapy efficacy at 8 Gy in our tumor model.

Keyphrases: Cancer, Cavitation, Nitrite, Radiotherapy, Microbubbles, Ultrasound, Provascular therapy, Hypoxia