This nanoelectroablation therapy effectively treats subdermal murine allograft tumors, autochthonous basal cell carcinoma (BCC) tumors in Ptch1+/−K14-Cre-ER p53 fl/fl mice, and UV-induced melanomas in C57/BL6 HGF/SF mice. Here we describe the first human trial of this modality. We treated 10 BCCs on three subjects with 100–1000 electric pulses 100 ns in duration, 30 kV/cm in amplitude, applied at 2 pulses per second. Seven of the 10 treated lesions were completely free of basaloid cells when biopsied and two partially regressed. Two of the 7 exhibited seborrheic keratosis in the absence of basaloid cells. One of the 10 treated lesions recurred by week 10 and histologically had the appearance of a squamous cell carcinoma. No scars were visible at the healed sites of any of the successfully ablated lesions. One hundred pulses were sufficient for complete ablation of BCCs with a single, one minute nanoelectroablation treatment.

We have identified an effective nanoelectroablation therapy for treating pancreatic carcinoma in a murine xenograft model. This therapy initiates apoptosis in a nonthermal manner by applying low energy electric pulses 100 ns long and 30 kV/cm in amplitude to the tumor. We first identified the minimum pulse number required for complete ablation by treating 30 tumors. We found that the minimum number of pulses required to ablate the tumor with a single treatment is between 250 and 500 pulses. We settled on a single application of either 500 or 1000 pulses to treat pancreatic carcinomas in 19 NIH-III mice. Seventeen of the 19 treated tumors exhibited complete regression without recurrence. Three mice died of unknown causes within 3 months after treatment but 16 lived for 270–302 days at which time we sacrificed them for histological analysis. In the 17 untreated controls, the tumor grew so large that we had to sacrifice all of them within 4 months.

The skeletal system plays an important role in the development and maturation process. Through the bone remodeling process, 10% of the skeletal system is renewed every year. Osteoblasts and osteoclasts are two major bone cells that are involved in the development of the skeletal system, and their activity is kept in balance. An imbalance between their activities can lead to diseases such as osteoporosis that are characterized by significant bone loss due to the overactivity of bone-resorbing osteoclasts. Our laboratory has developed a novel peptide, CK2.3, which works as both an anabolic and anti-resorptive agent to induce bone formation and prevent bone loss. We previously reported that CK2.3 mediated mineralization and osteoblast development through the SMAD, ERK, and AKT signaling pathways. In this study, we demonstrated the mechanism by which CK2.3 inhibits osteoclast development. We showed that the inhibition of MEK by the U0126 inhibitor rescued the osteoclast development of RAW264.7 induced by RANKL in a co-culture system with CK2.3. We observed that CK2.3 induced ERK activation and BMPRIa expression on Day 1 after stimulation with CK2.3. While CK2.3 was previously reported to induce the SMAD signaling pathway in osteoblast development, we did not observe any changes in SMAD activation in osteoclast development with CK2.3 stimulation. Understanding the mechanism by which CK2.3 inhibits osteoclast development will allow CK2.3 to be developed as a new treatment for osteoporosis.

DGAT2 is a transmembrane protein encoded by the DGAT2 gene that functions in lipid metabolism, triacylglycerol synthesis, and lipid droplet regulation. Cancer cells exhibit altered lipid metabolism and mutations in DGAT2 may contribute to this state. Using data from the Catalogue of Somatic Mutations in Cancer (COSMIC), we analyzed all cancer genetic DGAT2 alterations, including mutations, copy number variations and gene expression. We find that several DGAT2 mutations fall within the catalytic site of the enzyme. Using the Variant Effect Scoring Tool (VEST), we identify multiple mutations with a high likelihood of contributing to cellular transformation. We also found that D222V is a mutation hotspot neighboring a previously discovered Y223H mutation that causes Axonal Charcot-Marie-Tooth disease. Remarkably, Y223H has not been detected in cancers, suggesting that it is inhibitory to cancer progression. We also identify several single nucleotide polymorphisms (SNP) with high VEST scores, indicating that certain alleles in human populations have a pathogenic predisposition. Most mutations do not correlate with a change in gene expression, nor is gene expression dependent on high allele copy number. However, we did identify eight alleles with high expression levels, suggesting that at least in certain cases, the excess DGAT2 gene product is not inhibitory to cellular proliferation. This work uncovers unknown functions of DGAT2 in cancers and suggests that its role may be more complex than previously appreciated.

We have developed a low energy direct current pulsed electric field therapy for tissue ablation. This therapy applies 100 ns long electric pulses 30 kV/cm in amplitude using a contact electrode and triggers apoptosis in the treated tissue. Here we review the progress that has been made in understanding the mechanisms and targets of nanosecond pulsed electric fields (nsPEF) when applied to cells and tissues. This work began in 2001 in the laboratory of Karl Schoenbach who collaborated with biologists Stephen Beebe and Stephen Buescher to demonstrate the permeabilization of intracellular organelles. Since then over 100 papers have been published studying the cellular responses to nsPEF. We discuss these targets and cellular responses and introduce some new results from our group using nanoelectroablation to treat human pancreatic carcinoma in a murine xenograft model system. We have determined that 500 pulses 100 ns long and 30 kV/cm in amplitude are sufficient to ablate human pancreatic carcinomas growing in immunosuppressed mice and these ablated tumors do not recur for at least 300 days. We have also determined that the reactive oxygen species generation that is triggered within a minute after nsPEF treatment is Ca 2+ -dependent. In order bring this therapy into the clinic for the treatment of human tumors we are developing both a pulse generator as well as delivery electrodes to target the tumors to be treated. We describe the NanoBlate ® Model NB-1 100 ns pulse generator and the first human clinical trial data using nanoelectroablation to ablate basal cell carcinomas without scarring.