Muhammad Usman Ghani

Mutations in the CFTR gene play a pivotal role in the onset/severity of cystic fibrosis (CF). While our understanding of CFTR mutation classes is fragmented, this study focused on missense single nucleotide variants (SNVs) in the ABC transporter-like conserved site of the CFTR protein, employing various bioinformatics tools to identify deleterious amino acid substitutions (A.A.S). To gain a comparative understanding of the deleterious A.A.S in CFTR mutation classes, the in silico prediction of classified A.A.S by MutPred2 was cross-referenced with predictions of unclassified A.A.S. The study revealed twenty-five deleterious A.A.S in the conserved site. Nine of these (S549R, S549N, G551S, G551D, L558S, A559T, R560T, R560S, and A561E) were already classified. The in silico predictions for the remaining sixteen A.A.S exhibited similarities to molecular variations predicted for classified CFTR mutations and identified nine A.A.S falling under class II and seven A.A.S falling under class III, while four of these A.A.S in this conserved site may have effects of class II and III mutations. However, this classification is relative, warranting a comprehensive analysis to elucidate the intricacies of these nsSNVs. The combined use of modulators in therapy holds promise for more effective CF management, recognizing that CFTR mutations may exert effects that extend beyond a single class of mutation.

Cystic fibrosis (CF) is an inherited, autosomal recessive disorder that is caused by mutations in the gene encoding cystic fibrosis transmembrane conductance regulator (CFTR). CFTR maintains the balance between water and salts by transporting chloride ions along various epithelial surfaces. CFTR impairment affects the function of several organs, including the lungs. Newborn screening, prenatal diagnosis, and pharmacological interventions have altered the prevalence and incidence of cystic fibrosis. Although CFTR modulators are a promising treatment option, their ability to target and correct only one mutation at a time restricts their therapeutic potential. The development of genome editing technologies such as Clustered Regularly Interspaced Short Palindromic Repeats‐Cas(CRISPR‐Cas9) has the potential to correct genetic mutations, including those associated with CF, thereby offering a permanent treatment by fixing the root cause of CF. This article summarizes cystic fibrosis development, prognosis, and diagnosis, as well as possibilities for correcting various types of CFTR gene mutations. The review focuses on the potential of gene editing technologies to repair CFTR mutations and their applications in the advancement of CF treatment.

Background Pathogenic mutations in the CFTR gene disrupt the normal function of the chloride ion channel CFTR protein, resulting in Cystic Fibrosis (C.F.). Pakistan’s situation regarding C.F. mutation is largely unknown, complicating the disease management and treatment. This study is designed to identify the disease-causing CFTR mutations in the Pakistani C.F. cohort and perform an in silico analysis of rare/novel variants. Methodology Ninety-five C.F. patients were recruited from pediatric healthcare facilities in different regions of Pakistan. Initially, we investigated ∆F508 mutation in all patients, followed by whole exome sequencing (W.E.S.) of nineteen patients and in silico analysis of identified rare/novel mutations. Results and discussion Initial screening revealed that ∆F508 mutation was absent in 73.74% of cases. W.E.S. identified three novel variants (c.3036del/Q1012Hfs*11, c.488 A > T/p.K163M, c.2384del/S795Yfs*8), one rare variant (c.489 + 2T > C) previously reported in two Pakistani residing in U.K. and one (c.164 + 1G > T/p.?) extremely rare in other populations. Additionally, c.1705T > G/p.Y569D, c.653T > A/p.L218X, c.2125 C > T/p.R709X, and c.3484 C > T/p.R1162X were also identified. Most variants in our cohort are either frameshift or nonsense, while only two are missense variants. Alarmingly, most of these variants, except ∆F508, are non-responsive to modulator drugs, while the responsiveness of c.488 A > T/p.K163M is yet to be determined. High consanguinity (73.40%) and homozygous status of all mutations, except c.3036del/Q1012Hfs*11, are indicative of a high ratio of C.F. carriers in Pakistan. Conclusion These findings represent the diverse pattern of CFTR mutations within the Pakistani population, highlighting the imperative need to improve earlier C.F. diagnostic and management facilities and to conduct research on treatment strategies other than modulator therapies.

Thapsigargin, a sesquiterpene lactone derived from Thapsia garganica L., has demonstrated mixed potential as an anticancer agent due to its potent ability to disrupt calcium signaling and induce apoptosis. This review evaluates the chemopreventive and chemotherapeutic potential of thapsigargin, focusing on its molecular mechanisms and toxicity. An extensive literature review of studies published since 2015 was conducted using databases such as PubMed/MedLine and Science Direct. Findings indicate that thapsigargin’s primary mechanism is the inhibition of sarco/endoplasmic reticulum calcium ATPase, leading to endoplasmic reticulum stress and cell death in various cancer types. Despite these effects, thapsigargin’s non-specific cytotoxicity results in significant side effects, including organ damage and histamine-related reactions. Recent advances in targeted delivery, especially with the prodrug mipsagargin, initially suggested promise in minimizing these toxicities by selectively activating in cancer cells expressing prostate-specific membrane antigen (PSMA). However, the completion of clinical trials with no ongoing studies suggests that the viability of mipsagargin and other prodrugs remains uncertain, especially in light of the toxicities observed. While thapsigargin and its derivatives present a potential pathway in cancer treatment, their future role in oncology requires careful re-evaluation.

In the present study, Cinnamomum verum J. Presl. bark essential oil and its main component cinnamaldehyde was evaluated in vitro for neuraminidase (NA), transmembrane serine protease (TMPRSS2), and angiotensin converting enzyme 2 (ACE2) inhibitory activities. The chemical composition of C. verum essential oil was confirmed by both gas chromatography-mass spectrometry (GC/MS), and gas chromatography-flame ionization detection (GC-FID), where 75.9% (E)-cinnamaldehyde was the major component. The ACE2, NA, and TMPRSS2 enzyme inhibitions of C. verum bark essential oil at 20 μg/mL concentration, and (E)-cinnamaldehyde (5 μg/mL) were calculated and compared in the range of 54.2-89.9%. Molecular docking results supported that (E)-cinnam-aldehyde was specific to ACE2 with 89.9% inhibition. Our findings suggest further in vivo studies to confirm the effective and safe use of the essential oil as well as the (E)-cinnamaldehyde.

Circular RNAs (circRNAs) are a unique family of endogenous RNAs devoid of 3′ poly-A tails and 5′ end caps. These single-stranded circRNAs, found in the cytoplasm, are synthesized via back-splicing mechanisms, merging introns, exons, or both, resulting in covalently closed circular loops. They are profusely expressed across the eukaryotic transcriptome and offer heightened stability against exonuclease RNase R compared to linear RNA counterparts. This review endeavors to provide a comprehensive overview of circRNAs’ characteristics, biogenesis, and mechanisms of action. Furthermore, aimed to shed light on the potential of circRNAs as significant biomarkers in various cancer types. It has been performed an exhaustive literature review, drawing on recent studies and findings related to circRNA characteristics, synthesis, function, evaluation techniques, and their associations with oncogenesis. CircRNAs are intricately associated with tumor progression and development. Their multifaceted roles encompass gene regulation through the sponging of proteins and microRNAs, controlling transcription and splicing, interacting with RNA binding proteins (RBPs), and facilitating gene translation. Due to these varied roles, circRNAs have become a focal point in tumor pathology investigations, given their promising potential as both biomarkers and therapeutic agents. CircRNAs, due to their unique biogenesis and multifunctionality, hold immense promise in the realm of oncology. Their stability, widespread expression, and intricate involvement in gene regulation underscore their prospective utility as reliable biomarkers and therapeutic targets in cancer. As our understanding of circRNAs deepens, advanced techniques for their detection, evaluation, and manipulation will likely emerge. These advancements might catalyze the translation of circRNA-based diagnostics and therapeutics into clinical practice, potentially revolutionizing cancer care and prognosis.

Background: Alzheimer's disease (AD) is a neurodegenerative disorder characterized by extracellular amyloid β deposition and intercellular neurofibrillary tangles. AD is a complex disease with a strong genetic component, and it is classified into three phases by the Food and Drug Administration (US-FDA): pre-pathology, plaque formation, and neural destruction. A number of therapeutic agents have been developed to reduce AD pathology and improve cognition. Objectives: The objective of this paper was to review recent advancements in AD treatment and cure. Methodology: A literature search was conducted using the keywords "therapeutic developments for AD" and "natural products as a drug for AD." Publications related to AD were retrieved, and the NCBI Clinical trial database was also searched to check drug status. Results: Monoclonal antibodies against amyloid β plaques, tau protein, and microtubule-associated protein have shown positive results in animal models, but they have failed in clinical trials. This is likely due to the fact that these antibodies are designed to target specific proteins that are not always present in the human brain. Conclusion: Based on the findings of this review, it is suggested that medicines based on natural extracts and combination therapy would be least toxic and more effective than single therapy. This is because combination therapy targets multiple pathways involved in AD pathogenesis. There is also a dire need to initiate AD research in Pakistan and facilitate the Pakistani community in early diagnosis and cure.

Cystic fibrosis (CF) is a genetic monogenic disorder inherited in an autosomal recessive manner, marked by persistent airway infections in the endobronchial region. This condition leads to the gradual development of bronchiectasis and, ultimately, respiratory failure, emerging as the primary cause of mortality in individuals diagnosed with CF. Diagnosis is done depending on the patient’s symptoms and lung radiological findings like chest X‐rays and CTs. In younger patients and children, diagnosis becomes difficult due to overlapping symptoms with other diseases such as CF which is a rare genetic disease in our population. Diagnosis of CF usually relies on characteristic symptoms, a family history of CF, and an abnormal sweat chloride test, but in children, low sweat production during testing leads to false negative results. In this case report, a suspected patient with ambiguous respiratory symptoms underwent a comprehensive investigation revealing elevated CRP levels, TLC, and characteristic pulmonary manifestations on chest X‐ray, suggesting cystic fibrosis. Despite negative sweat chloride tests, the patient was analysed for potential candidate SNPs and was also tested for potential CFTR mutations to rule out CF, genetic analysis confirmed the diagnosis. Genetic testing plays a crucial role in diagnosing cystic fibrosis, even when traditional tests are inconclusive. Specific mutations like Δ508 deletion and rs213950 guide personalized treatment. Consanguinity and family history highlight genetic predisposition, while environmental factors may influence symptom onset. Further research is needed to understand these complexities and improve diagnostic and treatment approaches.

Cystic fibrosis (CF) is an autosomal recessive disorder caused by mutations in the CFTR gene. This study aimed to identify the spectrum of CFTR variants reported in individuals with CF from South Asia (ISA). We conducted a PubMed search for CFTR variants reported in ISA. Full text of original articles and case reports was read to compile data on reported variants. To gather additional data, we independently cross-referenced each variant with the CFTR Mutation Database and ClinVar. Our investigation identified a total of 92 CFTR variants reported across 30 articles. The most frequently tested, and reported variant was ΔF508 with a global frequency of 69.74%. Notably, we found 14 pathogenic CFTR mutations shared among ISA, originating from more than one South Asian country: ΔF508, 1525-1 G > A, G542X, S549N, R117H, S549R, R709X, V456A, Y569D, L1077P, 1161delC, 1898 + 1 G > T, G551D, and 2184insA. In summary, the higher prevalence of consanguinity and the limited availability of CF diagnostic resources in South Asia considerably contribute to the prevalence of genetic disorders like CF. The spectrum of CFTR mutations exhibits noticeable variations within South Asian and other populations. The inclusion of current study-enlisted CFTR gene variants is highly recommended for CF disease genetic testing in South Asia which may aid in achieving a precise diagnosis, enhancing disease management, and discovering drugs for currently untreatable genetic variants. It is also imperative to conduct a comprehensive study in this region, especially in previously unexplored countries such as Nepal, Bhutan, Maldives, and Bangladesh.