Targeted therapeutic strategies for treatment of HRAS mutant head and neck squamous cell carcinoma

RAS GTPases have three distinct family members: KRAS, NRAS and HRAS. It is well established that oncogenic RAS mutations in each of these RAS proteins play critical roles in tumor biology. Therefore, considerable effort has been made to understand the biology of RAS and its effectors in different cancer types. Both direct and indirect inhibitors of RAS are under clinical evaluation for cancer treatment. A challenge in the field is to understand how to overcome their limited success due to the complexity of RAS signaling and the emergence of resistance mechanisms. Head and neck squamous cell carcinoma (HNSCC) is one of the more common cancer types, impacting a significant number of patients globally. Despite other advances in the field of cancer therapeutics, the 5-year survival rate for HNSCC has not improved significantly. Both the disease and the current treatment strategies employed negatively impact patients' quality of life. Currently, the only targeted therapeutic agents approved for HNSCC are EGFR inhibitors and immune checkpoint inhibitors, limiting the available treatment options. One of the key limitations to designing new therapies against this tumor is insufficient information about actionable targets. HRAS is the most frequently mutated RAS oncogene present in HNSCC, and these mutations occur almost exclusively in the more devastating subset of HNSCC that are human papillomavirus (HPV) negative. My dissertation evaluated the role of HRAS as a tumor driver in HPV negative HNSCC and the efficacy of pharmacological inhibitors of HRAS membrane association and downstream signaling functions. In the first part of my dissertation, I established that HRAS is indeed a tumor driver in HRAS mutant HNSCC. I then investigated the target-based efficacy of the clinical candidate farnesyltransferase (FTI) tipifarnib for blocking HRAS membrane association and signaling, and its ability to impair proliferation and induce apoptosis in HRAS mutant HNSCC cell lines. I further performed CRISPR-Cas9 genetic loss-of-function screening to identify additional targets that could sensitize HNSCC to FTI treatment. Among the top hits, I found that combination of FTI with inhibitors of the RAS effectors ERK1/2 or PI3K enhanced efficacy and induced synergistic cell death. These could be potential therapeutic strategies for treatment of HRAS mutant HNSCC. In the second part of my doctoral work, I explored other combinations of targeted therapies that could be effective in HRAS mutant HNSCC. The RAF-MEK-ERK mitogen-activated protein kinase (MAPK) signaling cascade is one of the most well-established signaling pathways downstream of RAS. Inhibition of different nodes of this pathway has been exploited as a treatment strategy for RAS mutant tumors such as KRAS-mutant pancreatic cancer and NRAS-mutant melanoma. However, I found that HRAS mutant HNSCC were not sensitive to treatment with MAPK inhibitors alone. Inhibition of the MAPK pathway has been shown to cause upregulation of autophagy in KRAS mutant pancreatic cancer, creating a therapeutic vulnerability to autophagy inhibition. Co-treatment with ERKi and chloroquine, an autophagy inhibitor, induced apoptosis and tumor regression. I established that, similar to those tumors, ERK1/2 inhibition causes upregulation of autophagy in HRAS mutant HNSCC. However, synergistic cell death did not occur upon co-treatment with ERKi and chloroquine, highlighting differences between tumors harboring mutations in HRAS versus other RAS isoforms. Lastly, I characterized the global signaling changes induced upon ERK1/2 inhibition and identified signaling and epigenetic modulation mechanisms by which HRAS mutant HNSCC resist the effects of ERK1/2 inhibition. For example, inhibition of Aurora kinase and HDAC activity enhanced sensitivity to ERK inhibitors. My work identified potentially druggable targets that could sensitize HRAS mutant HNSCC cells to MAPK pathway inhibition.Doctor of Philosoph

Pre labour Rupture of Membranes at Term: Expectant Management vs Induction of labour

Objective: To determine the best approach to PROM at term. Study design: A randomized control trail. Place and duration of study: Obstetrics and Gynecology department of nishtar hospital from may 2017 to march 2018. Methodology: 100 women with PROM at term were selected. After consent, they were randomly assigned to expectant or induced group. Expectant group was managed conservatively while in the patients of the induced group induction of labour was done. All the data was entered on already designed Performa. Collected data was analyzed by SPSS software. Results: the duration between PROM to delivery was significantly reduced with a p value of <0.001 in induced group compared to the expectant group. Moreover, the difference of maternal morbidity and LSCS between induced and expectant group is not statistically significant. The difference observed in the neonatal outcomes was statistically significant showing higher rates of morbidity (asphyxia and sepsis) in the expectant group compared to the induced group. Conclusion:   induction of labour is the best approach towards the PROM at term compared to conservative management because of remarkable decrease in PROM to delivery interval and neonatal morbidity without significant rise of LSCS and maternal morbidity. Moreover, it gives the additional advantage of maternal satisfaction and decrease hospital stay time. Keywords: Pre Labour, Membranes Rupture, Induction

Comparison between bipolar diathermy and silk ligation technique during tonsillectomy

Objective: to compare the use of bipolar diathermy and silk ligation in tonsillectomy with respect to operative time and post op hemorrhage, in our setting. Study design: randomized controlled trail. Place and duration of study: nishtar hospital, multan from May 2016 to May 2018.  Methodology: After consent, 234 patients of chronic tonsillitis of 5 to 35 year of age were selected as per inclusion and exclusion criteria. They were randomly divided in to two groups by using lottery method. In group A, tonsillectomy was done and suture ligation was applied for hemostasis. In group B, tonsillectomy was done and bipolar diathermy was used for hemostasis. Data was entered and analyzed by using SPSS software. Results: In our study, mean age was 13.84 + 5.83 years. Majority of patients 55.13% were between 5-15 years of age. Out of 234 patients, 71.37% were male and 28.63% were females. The mean operative time in group A was 29.45 + 9.71 minutes and in group B was 16.37 + 4.38 minutes with p-value <0.0001. Secondary hemorrhage was seen in 2.56% patients in group A (suture ligation group) and 10.56% patients in group B (Bipolar diathermy group) with p-value of 0.016. Conclusion: bipolar diathermy method of hemostasis in tonsillectomy is fast but associated with statistically increase risk of secondary hemorrhage compared to silk ligation. Keywords: tonsillectomy, silk ligation, bipolar diathermy, operative interval, secondary hemorrhage

Coordinated Border Management (CBM): A Quantum Approach to Resolve Bilateral Conflicts between Pakistan and Afghanistan

This research work deals comprehensively with the chronology of the border issues at the Durand line. It focuses on the legitimacy of the Border line along with strong strategic analysis of the ongoing security situation of the region. In lieu of the prospects coordinated border management approach is offered with overview of the infrastructural plan and feasibility analysis for implementation in better interest of the neighboring states and regional security. In this research work, coordinated border management approach is supported in comparison to its operative forms internationally among union states and at the soft border points. It is not only suggestion for managing borders with different angles but a policy overview with estimated efficiency. This work will prove hallmark in the solution series for border management subjects

Genetic Engineering in Livestock: Zoology's Role in Improving Animal Health, Productivity, and Sustainability in Agriculture

This review paper provides an overview of the advancements and applications of genetic engineering techniques in livestock, providing an in-depth look at how genetic engineering has improved animal health, productivity, and sustainability in agriculture. In the introduction, genetic engineering is presented as an important tool for improving animal health, productivity, and sustainability in agriculture. Several genetic engineering tools and methods are discussed in the paper, including transgenic technology and genome editing techniques such as CRISPR-Cas9. A special focus is placed on the application of genetic engineering to livestock, specifically disease resistance and control, improved productivity, and environmental sustainability. Additionally, the review discusses the role of genetic engineering in improving animal health by introducing disease-resistant genes and eliminating disease-causing genes. Genetically engineered vaccines and enhanced biosecurity measures are also discussed in detail as well. A further examination of how genetic engineering can enhance livestock productivity is also included in the review, including the introduction of growth-promoting genes and the manipulation of metabolic pathways. As part of the program, nutritional content and allergenicity can also be enhanced in milk and meat products. As well as highlighting the importance of genetic engineering in promoting sustainable agriculture, the review also discusses how genetic engineering can reduce environmental impacts on agriculture. It is intended to reduce methane emissions and nitrogen and phosphorus waste, as well as optimize resources and land use through improved feed conversion efficiency and disease resistance in order to reduce the need for antibiotics. In this article, ethical and regulatory issues are discussed, including ethical concerns, current regulations, and guidelines for genetic engineering in livestock. At the conclusion of the review, future directions and challenges are discussed, emphasizing the need for continued research and responsible implementation in order to improve animal health, productivity, and sustainability in agriculture