Researchers have identified a molecule with enhanced antiproliferative activity in cancer cells
Vitamin D3 performs critical biological tasks such as maintaining bone mineral density, which reduces the risk of bone fracture. However, vitamin D3 is thought to have anti-cancer effect, since low levels of vitamin D3 and the concomitant overproduction of an enzyme called CYP24 have been linked to a bad prognosis for cancer patients. Molecules that restrict or inhibit CYP24 activity, as well as molecules that imitate the effect of vitamin D3, are now being studied as possible antiproliferative medicines for cancer treatment. However, many of the inhibitors and D3 analogs developed so far have had an insufficient clinical response as well as undesirable side effects. Madhu Biyani of Kanazawa University and colleagues have discovered a DNA-derived chemical that binds to and inhibits the action of CYP24 while also displaying promising antiproliferative activity. In addition, the research team provides extensive insights into the underlying molecular mechanisms at work.
The researchers tested a vast number of DNA aptamers, which are single-stranded DNA fragments with specialized three-dimensional structures that can bind to certain target molecules and have a functional effect when bound. They sought for DNA aptamers that bind to CYP24 but not to the similar enzyme CYP271B, which is responsible for vitamin D3 synthesis.
An initial longlist of 18 aptamer candidates was narrowed down to 11 candidates with specific molecular structures. The 11 representative aptamers were tested in vitro for their ability to inhibit CYP24. Four candidates that inhibited CYP24 but not CYP27B1 remained, one of which (Apt-7) were kept for further investigation
Biyani and colleagues performed simulations of Apt-7 binding to CYP24. A molecular docking scenario was obtained, which they checked experimentally by comparing the behavior of a mixture of vitamin D3 and CYP24 with and without Apt-7. The simulations and the experiments showed that Apt-7 results in the inhibition of CYP24 activity and that what happens is that the aptamer likely interferes with the enzyme’s active site. The researchers also performed high-speed atomic force microscopy on the binding of CYP24 and Apt-7 in real-time, confirming the molecular docking scenario obtained from simulations.
Finally, the research team studied the effect of Apt-7 at the cellular level by introducing the molecule to cancer cells. They observed significant CYP24 inhibition for a cancer cell line known to overexpress the CYP24 enzyme, thus showing antiproliferative activity. Quoting Biyani and colleagues, these findings “clearly characterized and proposed that a DNA aptamer-based molecule could be a promising lead candidate for anticancer therapy.
Reference: “Novel DNA Aptamer for CYP24A1 Inhibition with Enhanced Antiproliferative Activity in Cancer Cells” by Madhu Biyani, Kaori Yasuda, Yasuhiro Isogai, Yuki Okamoto, Wei Weilin, Noriyuki Kodera, Holger Flechsig, Toshiyuki Sakaki, Miki Nakajima and Manish Biyani, 18 April 2022, ACS Applied Materials & Interfaces.