Metallo-protease Peptidase M84 from Bacillusaltitudinis induces ROS-dependent apoptosis in ovarian cancer cells by targeting PAR-1.

We have purified Peptidase M84 from Bacillus altitudinis in an effort to isolate anticancer proteases from environmental microbial isolates. This metallo-protease had no discernible impact on normal cell survival, but it specifically induced apoptosis in ovarian cancer cells. PAR-1, a GPCR which is reported to be overexpressed in ovarian cancer cells, was identified as a target of Peptidase M84. We observed that Peptidase M84 induced PAR-1 overexpression along with activating its downstream signaling effectors NF-κB and MAPK to promote excessive reactive oxygen species (ROS) generation. This evoked apoptotic death of the ovarian cancer cells through the intrinsic route. In in vivo set-up, weekly intraperitoneal administration of Peptidase M84 in syngeneic mice significantly diminished ascites accumulation, increasing murine survival rates by 60%. Collectively, our findings suggested that Peptidase M84 triggered PAR-1-mediated oxidative stress to act as an apoptosis inducer. This established Peptidase M84 as a drug candidate for receptor mediated targeted-therapy of ovarian cancer.

Glycyrrhizin functionalized CuS Nanoprobes for NIR Light-based therapeutic mitigation of acne vulgaris.

Acne Vulgaris or Acne is a multifactorial bacterial infection caused by Propionibacterium acne, leading to inflammation and decreased quality of life, especially in adolescence. Currently, antibiotics and retinoids are preferred for treating acne. However, their continuous usage may lead to anti-microbial resistance and other side effects. Therefore, research on developing effective strategies to reduce antimicrobial resistance and improve acne healing is ongoing. The current work reports the synthesis and evaluation of near-infrared light-absorbing copper sulfide (CuS) nanoparticles loaded with a biomolecule, Glycyrrhizin (Ga). The photothermal efficacy studies, and in-vitro and in-vivo experiments indicated that the Ga-CuS NPs generated localized hyperthermia in acne-causing bacteria, leading to their complete growth inhibition. The results indicated that the Ga-Cus NPs possess excellent antibacterial and anti-inflammatory properties in the acne and inflammatory models. This could be from the synergistic effect of CuS NPs mediated mild Photothermal effect and inherent pharmacological properties of Ga. Further detailed studies of the formulations can pave the way for application in cosmetic clinics for the effective and minimally invasive management of Acne-like conditions.

Improved intra-operative decision making in RA-TKR with the help of pre-operative CT scan.

Background: Robotic-assisted total knee replacement (RA-TKR) is a significant advancement in orthopedic surgery, but intra-operative decision-making remains challenging. Pre-operative imaging techniques, particularly CT scans, have gained momentum, providing insights into the patient's anatomy, improving implant positioning and alignment. However, further research is needed to explore their influence on RA-TKR planning and execution. Materials and methods: The hospital based cross-sectional study was conducted in Orthopedics department of Sparsh Speciality Hospital, Bangalore & Sunshine Hospital, Hyderabad. A total of 1020 participants in the age group over 50 years during the study period were included based on convenient sampling. The axial CT images were taken preoperatively and RA-TKA was done for all the patients. Results: The study participant's average age was 64.01 ± 7.13. Out of 1020 patients 259 (24.4%) were males and 761 (74.6%) were females. The median femoral, tibia and Polyethylene predicted and the actual component were same with the side of surgery and BMI. The median femoral predicted actual component was significantly higher among the age category of more than 80 years when compared to other age groups. The median femoral, tibia and Polyethylene predicted was higher in males when compared to females. Conclusion: Pre-operative CT scans enhance RA-TKR procedures by providing precise anatomical insights, enhancing implant placement, and identifying potential issues, improving surgical outcomes and patient satisfaction.

CMT2A-linked MFN2 mutation, T206I promotes mitochondrial hyperfusion and predisposes cells towards mitophagy.

Mutations in Mitofusin2 (MFN2) associated with the pathology of the debilitating neuropathy Charcot-Marie-Tooth type 2A (CMT2A) are known to alter mitochondrial morphology. Previously, such mutations have been shown to elicit two diametrically opposite phenotypes - while some mutations have been causally linked to enhanced mitochondrial fragmentation, others have been shown to induce hyperfusion. Our study identifies one such MFN2 mutant, T206I that causes mitochondrial hyperfusion. Cells expressing this MFN2 mutant have elongated and interconnected mitochondria. T206I-MFN2 mutation in the GTPase domain increases MFN2 stability and renders cells susceptible to stress. We show that cells expressing T206I-MFN2 have a higher predisposition towards mitophagy under conditions of serum starvation. We also detect increased DRP1 recruitment onto the outer mitochondrial membrane, though the total DRP1 protein level remains unchanged. Here we have characterized a lesser studied CMT2A-linked MFN2 mutant to show that its presence affects mitochondrial morphology and homeostasis.

Cysteine-free cone snail venom peptides: Classification of precursor proteins and identification of mature peptides.

The cysteine-free acyclic peptides present in marine cone snail venom have been much less investigated than their disulfide bonded counterparts. Precursor protein sequences derived from transcriptomic data, together with mass spectrometric fragmentation patterns for peptides present in venom duct tissue extracts, permit the identification of mature peptides. Twelve distinct gene superfamiles have been identified with precursor lengths between 64 and 158 residues. In the case of Conus monile, three distinct mature peptides have been identified, arising from two distinct protein precursors. Mature acyclic peptides are often post-translationally modified, with C-terminus amidation, a feature characteristic of neuropeptides. In the present study, 20 acyclic peptides from Conus monile and Conus betulinus were identified. The common modifications of C-terminus amidation, gamma carboxylation of glutamic acid (E to ϒ), N-terminus conversion of Gln (Q) to a pyroglutamyl residue (Z), and hydroxylation of Pro (P) to Hyp (O) are observed in one or more peptides identified in this study. Proteolytic trimming of sequences by cleavage at the C-terminus of Asn (N) residues is established. The presence of an asparagine endopeptidase is strengthened by the identification of legumain-like sequences in the transcriptome assemblies from diverse Conus species. Such sequences may be expected to have a cleavage specificity at Asn-Xxx peptide bonds.

Identification and characterisation of emerging fish pathogens Aeromonas veronii and Aeromonas hydrophila isolated from naturally infected Channa punctata.

Naturally infected Channa punctata exhibiting bacterial septicemic syndrome including ulcerations along with mortality records were collected from a fish farm in Assam during winter season (early November 2020 to early January 2021). The moribund fishes were subjected for bacterial isolation followed by identification of the bacteria. Two dominant emerging bacterial pathogens were identified as Aeromonas veronii (isolate ZooGURD-01) and Aeromonas hydrophila (isolate ZooGURD-05) by standard biochemical characterization and 16S rRNA and rpo B gene amplification. Re-infection experiments of both the bacterial isolates in healthy disease-free C. punctata showed similar symptoms to that of natural infection thus confirming their virulence. The LD50 calculated during challenge test for both the isolates ZooGURD-01 and ZooGURD-05 found to be pathogenic at 2.6 × 104 and 1.6 × 104 CFU/fish respectively. Further PCR amplification of specific virulent genes (aerolysin, hemolysin and enterotoxin) confirmed pathogenicity for both isolates. Histopathological examinations of liver and kidney in re-infection experiments showed prominent changes supporting bacterial septicaemia. Antibiotic sensitivity pattern showed that the isolates ZooGURD-01 and ZooGURD-05 were sensitive to 22 and 19 out of 25 antimicrobials respectively. The present study was the first report on the mortality of farmed C. punctata associated with natural infection caused by A. veronii and A. hydrophila with no record of pathogenicity of A. veronii in C. punctata.

Relationship between femoral component placement and patient-specific anatomical rotational landmarks in robotic arm assisted total knee Arthroplasty- a multicentric study.

Introduction: Osteoarthritis of the knee is a common problem in the elderly, leading to severe morbidity. Total Knee Arthroplasty (TKA) is a widely validated surgery to provide a remarkable extent of knee function and simultaneously alleviates pain for knee osteoarthritis (OA). It is clearly understood that precision of the rotational alignment and accuracy of the technique in the placement of the femoral component is a prerequisite for excellent and successful outcomes of TKA. Advanced technology has now allowed surgeons to understand patient-specific variabilities in anatomical reference landmarks and the relationship of component positioning in relation to the reference landmarks to achieve accurate gap balancing with minimal soft tissue release.14 Robotic Arm Assisted-Total Knee Arthroplasty (RAA-TKA) is a semi-automated system that enables us in replicating the same. Using this technology, the bony resections, component positions, probable component sizing and gap balancing can be tentatively planned preoperatively with CT Scan Analysis and executed intraoperatively. Hence this study was undertaken to estimate the relationship between femoral component placement to normal rotational landmarks such as the Posterior Condylar Axis (PCA) and to quantitatively evaluate coronal and sagittal plane correction obtained. Also, we aimed to use the data to detect any anatomical variations in the study population and evaluate the accuracy of predicted component sizing, including gender-based evaluation. Materials and methods: A Prospective Observational Study of 1073 knees of patients of either sex above 50 years of age with Kellgren Lawrence Grade 4 Osteoarthritis of the knee which were confirmed with X-Ray undergoing RAA-TKA using MAKO Robotic System using Stryker Triathlon (Cruciate Substituting) CS Knee was conducted during the period between 2022 and 2023 in two South Indian hospitals specializing in joint replacement surgeries. Results: We found a statistically significant difference between the native Posterior Condylar Axis (PCA) (4.82 ± 2.15°) and final femoral component external rotation (3.24 ± 1.29°) with a p-value of <0.001 at 95% confidence interval. The accuracy of component size prediction was 99.8%. Also, analysis in our study has shown the most common implant sizes to be 4 in males and 2 in females. We also found no statistically significant difference based on age, size, laterality, or primary varus deformity. Conclusions: RAA-TKA provides patient-specific alignment/restricted kinematic alignment which might further enhance the outcome for the patient. Reliable deformity correction in coronal and sagittal planes can be achieved. Accurate flexion and extension gap balancing can be done through component placement and with minimal soft tissue dissection. Irrespective of all the advantages noted in RAA-TKA, further follow-up and long-term outcome studies are required to properly gauge and analyze this new technology.

Osteochondroma of Dorsal Scapula: A Case Report and Review of Literature.

Introduction: Osteochondroma of the scapula constitutes only 3-5% of all osteochondromas; osteochondroma on dorsal aspect of scapula is a rare entity. Diagnosis is almost always clinicoradiologically. Additional computed tomography scan and magnetic resonance imaging may be required for osteochondroma of flat bones such as scapula. Indications for surgery include pain, deformity, dysfunction, neural or vascular compromise, failure of conservative management, or in clinical settings with the high suspicion of malignant transformation and occasionally cosmesis. Outcome of a surgery should be assessed by Patient-Reported Outcome Measures (PROMs) which appraises what "matters to the patient." Case Report: A 10-year-old boy presented to us with painless swelling over the right upper back since 3 years of age and discomfort over the area while sleeping on his back for 6 months. Diagnosis confirmed it to be a pedunculated osteochondroma arising from the dorsal scapula. Here, we report the diagnosis, treatment, and successful Patient-Reported Outcome using QuickDASH© score for an osteochondroma of dorsal scapula using CARE© case reporting guidelines. Conclusion: We report a rare site of osteochondroma, review the relevant literature, and also stress upon the necessity of analyzing PROMs after surgical treatment of benign tumors of bone which would enable us to evaluate the result of surgery on symptoms, functioning, and health-related quality of life from the patient's perspective.

Can We Use Eplets (or Molecular) Mismatch Load Analysis to Improve Organ Allocation? The Hope and the Hype.

In recent years, there have been calls for implementation of "epitope matching" in deceased-donor organ allocation policies (later changed to "eplet matching"). Emerging data indeed support the use of molecular mismatch load analysis in specific patient groups, with the objective of posttransplant stratification into different treatment arms. For this purpose, the expectation is to statistically categorize patients as low- or high-immune-risk. Importantly, these patients will continue to be monitored' and their risk category, as well as their management, can be adjusted according to on-going findings. However, when discussing deceased donor organ allocation and matching algorithms, where the decision is not modifiable and has lasting impact on outcomes, the situation is fundamentally different. The goal of changing allocation schemes is to achieve the best possible HLA compatibility between donor and recipient. Immunologically speaking, this is a very different objective. For this purpose, the specific interplay of immunogenicity between the donor and any potential recipient must be understood. In seeking compatibility, the aim is not to redefine matching but to identify those mismatches that are "permissible" or' in other words, less immunogenic. In our eagerness to improve transplant outcome, unfortunately, we have conflated the hype with the hope. Terminology is used improperly, and new terms are created in the process with no sufficient support. Here, we call for a cautious evaluation of baseline assumptions and a critical review of the evidence to minimize unintended consequences.

Facets-Directed Epitaxially Grown Lead Halide Perovskite-Sulfobromide Nanocrystal Heterostructures and Their Improved Photocatalytic Activity.

Lead halide perovskite nanocrystal heterostructures have been extensively studied in the recent past for improving their photogenerated charge carriers mobility. However, most of such heterostructures are formed with random connections without having strong evidence of epitaxial relation. Perovskite-chalcohalides are the first in this category, where all-inorganic heterostructures are formed with epitaxial growth. Going beyond one facet, herein, different polyhedral nanocrystals of CsPbBr3 are explored for facet-selective secondary epitaxial sulfobromide growths. Following a decoupled synthesis process, the heterojunctions are selectively established along {110} as well as {200} facets of 26-faceted rhombicuboctahedrons, the {110} facets of armed hexapods, and the {002} facets of 12-faceted dodecahedron nanocrystals of orthorhombic CsPbBr3. Lattice matching induced these epitaxial growths, and their heterojunctions have been extensively studied with electron microscopic imaging. Unfortunately, these heterostructures did not retain the intense host emission because of their indirect band structures, but such combinations are found to be ideal for promoting photocatalytic CO2 reduction. The pseudo-Type-II combination helped here in the successful movement of charge carriers and also improved the rate of catalysis. These results suggest that facet-selective all-inorganic perovskite heterostructures can be epitaxially grown and this could help in improving their catalytic activities.

CMT2A-linked mitochondrial hyperfusion-driving mutant MFN2 perturbs ER-mitochondrial associations and Ca2+ homeostasis.

BACKGROUND INFORMATION: Mitofusin2 (MFN2), an important molecular player that regulates mitochondrial fusion, also helps maintain the inter-organellar contact sites, referred as mitochondria associated membranes (MAMs) that exist between the ER and mitochondria. The study deals with a mutant of MFN2, R364W-MFN2, linked with the neuropathy, Charcot Marie Tooth (CMT) disease. Previous studies show that this mutant promotes mitochondrial hyperfusion. Here, we try to decipher the role of R364W-MFN2 in affecting the ER mitochondrial associations at the MAM junctions and inter-organellar calcium signalling between the ER and the mitochondria. RESULTS: Our results show that R364W-MFN2 altered ER-mitochondria association at the MAM junctions, predisposed mitochondria towards cellular stress with the mitochondria undergoing rapid fission upon induction of mild stress and perturbs inter-organellar calcium homeostasis. CONCLUSION: The results indicate that R364W-MFN2 not only affects mitochondrial morphology and dynamics but also modulate its interaction with the ER and Ca2+ signalling between the two organelles. SIGNIFICANCE: This study provides significant insight that presence of the R364W-MFN2 mutation makes cells susceptible towards stress, thus negatively affecting cellular health which altogether might culminate in the form of the CMT neuropathy.

Tuning Crystal Plane Orientation in Multijunction and Hexagonal Single Crystalline CsPbBr3 Perovskite Disc Nanocrystals.

Two-dimensional-shaped CsPbBr3 platelet nanocrystals are widely studied for their bright high energy emission and self-assembly. These nanostructures are in orthorhombic phase, have a square shape, and have the vertical axis [001] perpendicular to the basal plane. Moreover, these are mostly single-crystalline structures with a continuous lattice and appear like slices of cube nanocrystals. In contrast, herein, multijunction and hexagonal single crystalline 2D discs of CsPbBr3 are reported to have all their vertical axes [100]. These are obtained by using the perovskite derivative of tetragonal Cs3MnBr5 as the parent material and subsequent B-site Pb(II) introduction in the presence of phenacyl bromide at different reaction temperatures. At low temperature, multijunction discs having random orientations of two horizontal axes [010] and [001] from one to another segment are observed. Orientations of planes remained random as both coherent and incoherent twin planes were observed at their boundaries. However, the number of junctions/segments was reduced at higher temperature, and finally hexagonal single crystalline discs remained as the ultimate product. Analysis suggested that the crystal nature of parent Cs3MnBr5 and temperature-dependent variation in the rate of Pb(II) insertions determined the nature of discs having randomly oriented or static planes in the entire nanostructure. Not only in 2D discs but also, 3D nanocrystals having similar segments with different orientations are formed upon Pb(II) exchange with Mn(II) alloyed cubic CsBr. Hexagonal single crystalline and segmented multijunction CsPbBr3 discs remain unique among 2D perovskites nanostructures, and their formation mechanism indeed introduced new fundamentals of the crystallization process of these emerging energy materials.

Evaluation of anterior approach in failed closed reduction and delayed presentation of supracondylar humerus fractures in children.

BACKGROUND: The treatment of displaced paediatric supracondylar fracture is a challenging problem and requires strict vigilance and a proper management protocol. Prospective investigation of the treatment options for fractures that cannot be reduced by closed reduction is recommended in literature. Operative treatment is indicated for the fractures that cannot be reduced satisfactorily by closed methods. It is also considered the best option for late presenting fractures. The aim of this study was to assess and compare the clinical outcome using open reduction through anterior approach in delayed presentation and failed closed reduction of supracondylar fracture humerus in children. MATERIALS AND METHODS: 15 patients of failed closed reduction and 11 patients of delayed presentation of supracondylar humerus fractures were operated with anterior approach. The demographic data, time from injury to presentation and from admission to surgery, reasons for delayed presentation, type of fracture, operative findings and time, K-wire configuration, length of hospitalization, post operative complications were noted. The patients were followed up for a period of 12 months and final range of motion, Baumann's angle, and cosmetic, functional and overall outcome by Flynn's criteria were evaluated and analyzed. RESULTS: The overall outcome was very satisfactory according to Flynn's criteria. 80.77% patients had excellent, 15.38% patients had good, and 3.85% patient had fair results with no poor results. Our results show distinct advantage of anterior approach which are on a par with or better than the previous studies using anterior approach, adding to their evidence. CONCLUSION: Open reduction using anterior approach is a very safe, logical and effective technique of treating failed closed reduction or late presentation of supracondylar fractures humerus in children with excellent cosmetic and functional results, and offers distinctive advantage over other approaches.

Septic Arthritis in Pediatric Hemophilia A: A Case Report and Review of Literature.

Hemophilia A is a hereditary bleeding disorder due to coagulation factor VIII deficiency. Septic arthritis is in itself a dreaded infective condition of the joint. It is much more common in hemophilia patients than the general population and is associated with considerably higher morbidity. It presents as a difficult clinical and therapeutic problem, necessitating prompt diagnosis and intensive therapy. There was under-diagnosis of blood coagulation disorders in developing countries like India which has improved in the present time. But there is still considerable uncertainty about proper management guidelines, protocols and resources settings for treating this complication in hemophilia patients. Hereby we present a case of a 9-year-old hemophilia A patient with septic arthritis of knee joint treated at a tertiary hospital centre with available present recommended guidelines in literature, and stress upon the necessity of further formulating and improvising management guidelines for treatment and prevention of this complication in Hemophilia.

Molecular histocompatibility beyond Tears: The next generation version.

Human Leukocyte Antigens (HLA) matching at the serological level used to serve as the measure of histocompatibility between organ donors and recipients. With advancements in HLA typing methods more precise HLA mismatch assessment tools were developed to measure dissimilarities at the molecular level, collectively referred to as Molecular Mismatch load analysis tools. Currently, several software are aimed at deciphering the dissimilarities using somewhat different immunologic rationales. Our goal, in this review is to provide a basic overview of the different computational approaches, provide clinical cases to contextualize concerns regarding the lack of assessment of immunogenicity, and present our personal view regarding the gaps and the needs of our field.

MITOL-mediated DRP1 ubiquitylation and degradation promotes mitochondrial hyperfusion in a CMT2A-linked MFN2 mutant.

Mutations in mitofusin 2 (MFN2) that are associated with the pathology of the debilitating neuropathy Charcot-Marie-Tooth type 2A (CMT2A) are known to alter mitochondrial morphology. One such abundant MFN2 mutation, R364W, results in the generation of elongated, interconnected mitochondria. However, the mechanism leading to this mitochondrial aberration remains poorly understood. Here, we show that mitochondrial hyperfusion in the presence of R364W-MFN2 is due to increased degradation of DRP1 (also known as DNM1L). The E3 ubiquitin ligase MITOL (also known as MARCHF5) is known to ubiquitylate both MFN2 and DRP1. Interaction with and subsequent ubiquitylation by MITOL is stronger in the presence of wild-type MFN2 than with R364W-MFN2. This differential interaction of MITOL with MFN2 in the presence of R364W-MFN2 renders the ligase more available for DRP1 ubiquitylation. Multi-monoubiquitylation and proteasomal degradation of DRP1 in R364W-MFN2 cells in the presence of MITOL eventually leads to mitochondrial hyperfusion. Here, we provide a mechanistic insight into mitochondrial hyperfusion, while also reporting that MFN2 can indirectly modulate DRP1 - an effect not shown previously. This article has an associated First Person interview with the first author of the paper.

Lipidomic Profiling of Clinical Prostate Cancer Reveals Targetable Alterations in Membrane Lipid Composition.

Dysregulated lipid metabolism is a prominent feature of prostate cancer that is driven by androgen receptor (AR) signaling. Here we used quantitative mass spectrometry to define the "lipidome" in prostate tumors with matched benign tissues (n = 21), independent unmatched tissues (n = 47), and primary prostate explants cultured with the clinical AR antagonist enzalutamide (n = 43). Significant differences in lipid composition were detected and spatially visualized in tumors compared with matched benign samples. Notably, tumors featured higher proportions of monounsaturated lipids overall and elongated fatty acid chains in phosphatidylinositol and phosphatidylserine lipids. Significant associations between lipid profile and malignancy were validated in unmatched samples, and phospholipid composition was characteristically altered in patient tissues that responded to AR inhibition. Importantly, targeting tumor-related lipid features via inhibition of acetyl-CoA carboxylase 1 significantly reduced cellular proliferation and induced apoptosis in tissue explants. This characterization of the prostate cancer lipidome in clinical tissues reveals enhanced fatty acid synthesis, elongation, and desaturation as tumor-defining features, with potential for therapeutic targeting. SIGNIFICANCE: This study identifies malignancy and treatment-associated changes in lipid composition of clinical prostate cancer tissues, suggesting that mediators of these lipidomic changes could be targeted using existing metabolic agents.

Case Report of a Rare Cause of Reactive Arthritis: Leptospirosis.

INTRODUCTION: Leptospirosis is a zoonosis caused by infection with pathogenic Leptospira species. Leptospirosis has protean manifestations and rare, unusual presentations should be kept in mind in relevant epidemiological scenario. Reactive arthritis refers to acute non-purulent arthritis complicating an infection elsewhere in the body. It is attributed to an immune activation following certain infections; it is, therefore considered as aseptic arthritis. Very few case reports are available attributing leptospirosis as an established cause of reactive arthritis. We present a case of reactive arthritis of the hip joint due to leptospirosis. CASE REPORT: Here, we present a case of a 12-years- old female child who was admitted to our hospital with complaints of fever, headache, and pain in the right hip joint since past 5 days from admission. Subsequent elaboration revealed a past history of fever, headache, and myalgia for around 5-7 days around a week before the present complaints. There was rat infestation near her house and her father was working as sewage cleaner. Routine investigations, Ultrasonography (USG), Magnetic Resonance Imaging (MRI) of both hips and subsequently, diagnostic hip aspiration was performed. USG revealed synovitis, MRI revealed hip joint arthritis of infective or inflammatory origin. Diagnostic hip aspiration was negative for any microorganism. On 10th day of admission, patient started developing icterus with yellowish discolouration of urine. Patient was evaluated for the cause of jaundice. Screening for Leptospira was positive. Synovial biopsy of hip was performed, which showed inflammation with no specific pathology and no growth of any microorganism. In addition, Leptospira IgM MAC ELISA was done which was positive. Patient was thus confirmed to be having leptospirosis and reactive arthritis as a consequence of it. CONCLUSION: The presentation of reactive arthritis secondary to leptospirosis is rare. Leptospirosis can be an etiological factor for reactive arthritis, especially if reactive arthritis is complicated with jaundice.

An integrated functional and clinical genomics approach reveals genes driving aggressive metastatic prostate cancer.

Genomic sequencing of thousands of tumors has revealed many genes associated with specific types of cancer. Similarly, large scale CRISPR functional genomics efforts have mapped genes required for cancer cell proliferation or survival in hundreds of cell lines. Despite this, for specific disease subtypes, such as metastatic prostate cancer, there are likely a number of undiscovered tumor specific driver genes that may represent potential drug targets. To identify such genetic dependencies, we performed genome-scale CRISPRi screens in metastatic prostate cancer models. We then created a pipeline in which we integrated pan-cancer functional genomics data with our metastatic prostate cancer functional and clinical genomics data to identify genes that can drive aggressive prostate cancer phenotypes. Our integrative analysis of these data reveals known prostate cancer specific driver genes, such as AR and HOXB13, as well as a number of top hits that are poorly characterized. In this study we highlight the strength of an integrated clinical and functional genomics pipeline and focus on two top hit genes, KIF4A and WDR62. We demonstrate that both KIF4A and WDR62 drive aggressive prostate cancer phenotypes in vitro and in vivo in multiple models, irrespective of AR-status, and are also associated with poor patient outcome.