Biomedicine & Translational Research
The cellular molecules are the basis of life and the mechanistic tools for scientists to study and understand how the living system works. We utilize various cellular and molecular biology techniques and integrate multidisciplinary principles involving biochemistry, cell biology, immunology, pathology, and pharmacology to explore and understand the normal physiology and development of human diseases like cancer.
In collaboration with National Cancer Hospital and Research Center (NCHRC) and exploiting patients’ clinical and molecular data, we aim to identify and establish different diagnostic, prognostic and treatment biomarkers for different types of cancer. We also aim to explore the potential of local natural products in treating various cancers.
Synthesis and characterization of non-viral vectors for efficient gene delivery.
Viral vectors are usually used for gene therapy to deliver genes into cells. However, the delivery of genes by virus raises safety concerns. As an alternative, synthetic polymer-based (non-viral) vectors have emerged as a promising gene vehicle because cationic polymers have the ability to make complexation with genes to form polymeric nanoparticles for gene transfection. Importantly, a successful gene delivery requires to overcome several limitations, such as cytotoxicity, cellular uptake and endosomal escape. Although each barrier represents a critical step in determining the final efficiency of gene delivery, the release of DNA from endosomes, that is, endosomal escape prior to their degradation in lysosomes, appears to be a major bottleneck. It is now well-documented that polyethylenimine (PEI) induces endosomal escape due to its buffering capacity. Although the high density of positive charges of PEI is rewarding to polymer for buffering capacity, it contributes to high cytotoxicity. Hence, much attention has been given to design polymeric vectors aiming for efficient transfection with low cytotoxicity. Although the scope of non-viral gene delivery is promising, no report has been published from any institute or University till date from Nepal. This study aims to synthesize various gene delivery vectors, and test them for possible use in gene transfection.
Utilizing cationic peptides for the development of novel antibacterial hydrogels for addressing bacterial infections in an environment of increasing bacterial resistance to existing antibiotics.
Hydrogels are becoming popular in the field of biomedical research due its applicability in many functional applications. While conventional hydrogels use synthetic polymers, hydrogels formed by molecules that self-assemble via noncovalent forces are gaining popularity due to enhanced biocompatibility. Therefore Low Molecular Weight (LMW) based hydrogelators are an attractive alternative to the synthetic polymer gelators due to its reduced cost and synthetic customizability. My previous work has shown that cationic peptides such as (FKFK)2 and Fmoc-Phe derivatives have the ability to spontaneously self-assemble to form rigid hydrogels that satisfies most of the criteria needed for cell culture applications. It has also been reported that cationic polymers and peptides have the ability to display efficient antibacterial activities. Hence, we can hypothesize that the polymeric fibrils of these cationic molecules can be also utilized to form hydrogels that can support mammalian cells while displaying antibacterial properties. Such gels are highly desirable because these gels can be potentially applied against bacterial infectivity in countries such as Nepal where bacterial infections impose a great threat in the general public health. For our research, we plan to utilize these gels to probe its ability to sustain mammalian cells while inhibiting bacterial cells by using spectrophotometer, fluorescence/confocal microscope to quantify live/dead assay of the cells within the gel.
Monitoring circulating Cell free DNA and KRAS mutation presence in Circulation as a candidate biomarker in colorectal cancer patients of Nepal
Colorectal cancer (CRC) originates in cells of the colon or rectum. CRC is the third most common cancer worldwide. Surgery, radiation and chemotherapy remained the mainstream care of treatment. CRC is one of the major disease problems in Nepal. Patients not only die of loco-regional disease but also due the recurrent disease. There is an urgent need for biomarkers that allow early detection of relapse in colorectal cancer. We here propose that one such biomarker may be Circulating Cell-free DNA (CCFDNA) measurement. Recently, circulating cell-free DNA has attracted considerable interest in the cancer biomarker field. Increased levels of CCFDNA has been observed in cancer patients as compared to healthy donors. In a developing country like Nepal, finding a cancer biomarker using high throughput technology is challenging due to financial and human resources reasons. In addition to that, biomarkers should be easily isolated and should have a clinical utility. CCFDNA can easily be isolated from the plasma of cancer patients indicating a promising candidate as a non-invasive biomarker of cancer. Measurement of CCFDNA over time may allow oncologists to detect and treat recurrence early enough to improve disease outcomes. However, it has not been evaluated whether monitoring CCFDNA has any diagnostic, predictive /prognostic utility in context of colorectal cancer patients from Nepal. In addition, KRAS mutation is one of the most frequent molecular abnormalities in colorectal cancer. KRAS mutation presence in CCFDNA associates with shorter survival of CRC patients. This study will also evaluate KRAS mutation in CRC using CCFDNA.
Exploration of Alternative Vectors of Leishmania Parasites at Higher Altitudes of Nepal.
Nepal Health Research Council (NHRC)
The main aim of this research is explore and understand the distribution of the sand fly (Phlebotomus argentipes) as well as the potential alternative vector capable of carrying Leishmania parasites in the high altitude regions of Nepal with particular focus in the Dolpa district since several cases have been reported from the area either by our team or others.
Utilization of potent tripeptide for the development of novel anti-inflammatory systems towards inflammatory bowel disease treatment
The occurrence of inflammatory bowel disease (IBD) has experienced an increase worldwide, including developing countries like Nepal. However, current therapies mainly provide symptomatic relief and are often associated with side effects. Parenterally delivered monoclonal antibodies against TNF-α act as second line treatment but it is also linked with life-threatening side effects. The proposed project aims to help in developing a safer, effective and cheaper IBD therapy by using a potent tripeptide, lysine proline-valine (KPV). However, KPV is a small hydrophilic molecule, which is rapidly eliminated after oral delivery, and is also tricky to load into a formulation. To overcome these challenges, we propose modification of KPV with a cell penetrating molecule, lauryl-octaarginine, which would increase its size and hydrophobicity and form nanocomplex with poly-γ-glutamic acid. Thus, we hypothesize that the nanocomplexes formed from modified KPV can show its anti-inflammatory properties after oral administration. Other benefits of this system are biocompatibility, biodegradability and the use of patient friendly oral routes. Moreover, such novel systems are very desirable as KPV has no notable side effects compared to other treatment lines. Hence, such systems can have a potential use against IBD and other inflammatory conditions in poor countries, such as Nepal, where chronic socioeconomic impact of such disease impose a great threat to the general public health.