Disrupted intercellular bridges and spermatogenesis in fatty acyl-CoA reductase 1 knockout mice: A new model of ether lipid deficiency.

Peroxisomal fatty acyl-CoA reductase 1 (FAR1) is a rate-limiting enzyme for ether lipid (EL) synthesis. Gene mutations in FAR1 cause a rare human disease. Furthermore, altered EL homeostasis has also been associated with various prevalent human diseases. Despite their importance in human health, the exact cellular functions of FAR1 and EL are not well-understood. Here, we report the generation and initial characterization of the first Far1 knockout (KO) mouse model. Far1 KO mice were subviable and displayed growth retardation. The adult KO male mice had smaller testes and were infertile. H&E and immunofluorescent staining showed fewer germ cells in seminiferous tubules. Round spermatids were present but no elongated spermatids or spermatozoa were observed, suggesting a spermatogenesis arrest at this stage. Large multi-nucleated giant cells (MGC) were found lining the lumen of seminiferous tubules with many of them undergoing apoptosis. The immunofluorescent signal of TEX14, an essential component of intercellular bridges (ICB) between developing germ cells, was greatly reduced and mislocalized in KO testis, suggesting the disrupted ICBs as an underlying cause of MGC formation. Integrative analysis of our total testis RNA-sequencing results and published single-cell RNA-sequencing data unveiled cell type-specific molecular alterations underlying the spermatogenesis arrest. Many genes essential for late germ cell development showed dramatic downregulation, whereas genes essential for extracellular matrix dynamics and cell-cell interactions were among the most upregulated genes. Together, this work identified the cell type-specific requirement of ELs in spermatogenesis and suggested a critical role of Far1/ELs in the formation/maintenance of ICB during meiosis.

AI-2 quorum sensing-induced galactose metabolism activation in Streptococcus suis enhances capsular polysaccharide-associated virulence.

Bacteria utilize intercellular communication to orchestrate essential cellular processes, adapt to environmental changes, develop antibiotic tolerance, and enhance virulence. This communication, known as quorum sensing (QS), is mediated by the exchange of small signalling molecules called autoinducers. AI-2 QS, regulated by the metabolic enzyme LuxS (S-ribosylhomocysteine lyase), acts as a universal intercellular communication mechanism across gram-positive and gram-negative bacteria and is crucial for diverse bacterial processes. In this study, we demonstrated that in Streptococcus suis (S. suis), a notable zoonotic pathogen, AI-2 QS enhances galactose utilization, upregulates the Leloir pathway for capsular polysaccharide (CPS) precursor production, and boosts CPS synthesis, leading to increased resistance to macrophage phagocytosis. Additionally, our molecular docking and dynamics simulations suggest that, similar to S. pneumoniae, FruA, a fructose-specific phosphoenolpyruvate phosphotransferase system prevalent in gram-positive pathogens, may also function as an AI-2 membrane surface receptor in S. suis. In conclusion, our study demonstrated the significance of AI-2 in the synthesis of galactose metabolism-dependent CPS in S. suis. Additionally, we conducted a preliminary analysis of the potential role of FruA as a membrane surface receptor for S. suis AI-2.

Recent advances of Pin1 inhibitors as potential anticancer agents.

Pin1 (proline isomerase peptidyl-prolyl isomerase NIMA-interacting-1), as a member of PPIase family, catalyzes cis-trans isomerization of pThr/Ser-Pro amide bonds of its substrate proteins, further regulating cell proliferation, division, apoptosis, and transformation. Pin1 is overexpressed in various cancers and is positively correlated with tumor initiation and progression. Pin1 inhibition can effectively reduce tumor growth and cancer stem cell expansion, block metastatic spread, and restore chemosensitivity, suggesting that targeting Pin1 may be an effective strategy for cancer treatment. Considering the promising therapeutic effects of Pin1 inhibitors on cancers, we herein are intended to comprehensively summarize the reported Pin1 inhibitors, mainly highlighting their structures, biological functions and binding modes, in hope of providing a reference for the future drug discovery.

Percutaneous transforaminal endoscopic discectomy for Upper Lumbar Disc Herniation versus lower lumbar disc herniation: clinical outcomes and technical consideration.

BACKGROUND: Upper lumbar disc herniation (ULDH) accounts for 1-10% of all lumbar disc herniations (LDH). This study aimed to evaluate the clinical characteristics and outcomes of patients with ULDH who underwent percutaneous transforaminal endoscopic discectomy (PTED) compared with those with lower LDH. METHODS: 60 patients with ULDH or L4-L5 LDH treated with PTED between May 2016 and October 2021. MacNab criteria, visual analog scale (VAS) of back pain and leg pain, and Japanese Orthopedic Association (JOA) were evaluated before and after surgery. RESULTS: In the L1-L3 group, 59.1% of the patients had a positive femoral nerve tension test, and 81.8% of the patients had a sensory deficit. Both groups showed significant improvements in VAS scores for low back and leg pain, and JOA scores postoperatively. No significant differences in the degree of improvement were observed between the two groups. The excellent/good rate was 81.8% in the L1-L3 group and 84.2% in the L4-L5 group, showing no significant difference. CONCLUSION: PTED has comparable efficacy in treating ULDH as it does in treating lower LDH, it is a safe and effective treatment method for ULDH.

Sorting for secreted molecule production using a biosensor-in-microdroplet approach.

Sorting large libraries of cells for improved small molecule secretion is throughput limited. Here, we combine producer/secretor cell libraries with whole-cell biosensors using a microfluidic-based screening workflow. This approach enables a mix-and-match capability using off-the-shelf biosensors through either coencapsulation or pico-injection. We demonstrate the cell type and library agnostic nature of this workflow by utilizing single-guide RNA, transposon, and ethyl-methyl sulfonate mutagenesis libraries across three distinct microbes (Escherichia coli, Saccharomyces cerevisiae, and Yarrowia lipolytica), biosensors from two organisms (E. coli and S. cerevisiae), and three products (triacetic acid lactone, naringenin, and L-DOPA) to identify targets improving production/secretion.

Rare Sagittal Compensatory Balance in Neuromuscular Scoliosis Complicated by Hydrocephalus: A Case Report.

Objective: This case report aims to present a rare case of thoracic lordosis and lumbar kyphosis and describe the posterior instrumented scoliosis correction performed. Case presentation: A 59-year-old female presented with low back pain. She had undergone ventriculoperitoneal shunt placement 8 years ago. I scored 76 on the Wechsler Adult Intelligence Scale. MRI of the lumbar spine showed spinal canal stenosis at L3/4, L4/5, and L5/S1. Full spine X-ray revealed thoracic lordosis and lumbar kyphoscoliosis， the coronal imbalance, and the sagittal compensatory balance. In order to avoid the risk of brain swelling and paraplegia, pedicle subtraction osteotomies (PSO) in the L2 and lumbar posterior instrumented scoliosis correction were performed under electroencephalogram and neuro electrophysiological monitoring. Shoulder imbalance was observed 1 year after surgery, but there was no loss of lumbar correction. Conclusion: In future cases of complex spinal deformity, it is important to observe whether there is cerebral ventricular dilatation on MRI before the operation. If severe thoracic lordosis is combined with lumbar scoliosis, over-correcting the lumbar scoliosis should be avoided to prevent shoulder imbalance.

Clinical outcome of percutaneous endoscopic lumbar decompression in treatment of elderly patients with lumbar spinal stenosis: a matched retrospective study.

PURPOSE: This retrospective cohort study aimed to evaluate the efficacy and safety of percutaneous endoscopic lumbar decompression (PELD) in elderly patients with lumbar spinal stenosis (LSS). STUDY DESIGN: A matched retrospective study. SETTING: The research was conducted in Beijing Chaoyang Hospital, Capital Medical University, China. METHODS: This study included patients treated with PELD for LSS from September 2016 to September 2020. Patients with LSS aged ≥ 80 years were screened according to the inclusion and exclusion criteria as the study group, and then the same number of patients with LSS aged 50-80 years were matched according to gender, stenosis type, and surgical segment as the control group. Preoperative patient status was assessed using the Charlson comorbidity index (CCI) and the American Society of Anesthesiologists (ASA) physical status classification score. Clinical outcomes were assessed using the visual analog scale (VAS), Oswestry Disability Index (ODI) scores, modified Macnab criteria, radiological parameters and complication rates. RESULTS: A total of 624 LSS patients met the screening criteria between September 2016 and September 2020, with 47 LSS patients ≥ 80 years old serving as the study group. Forty-seven LSS patients aged 50-80 years were matched to the study group according to gender, stenosis type, and stenosis segment. The CCI score (1.77 ± 1.67) and ASA classification (2.62 ± 0.74) of the study group were significantly higher than the CCI score (0.66 ± 0.96) and ASA classification (1.28 ± 0.54) of the control group, and the difference was statistically significant. Compared with preoperative data, postoperative ODI, leg pain VAS scores and back pain VAS scores were significantly improved in both groups (p < 0.05). However, no significant difference was found between two groups in preoperative and postoperative ODI, leg pain VAS scores and back pain VAS scores (p > 0.05). The operation time and postoperative hospital stay in control group were significantly lower than those in study (p < 0.05), but there was no significant difference in blood loss between the two groups (p > 0.05). Besides, overall radiological parameters were comparable in elder and younger patients (p > 0.05), and disc height (DH), lumbar lordosis and segmental lordosis decreased after two year follow-up in both groups (p < 0.05). In addition, complication rates were similar between the two groups (p > 0.05), and no serious complications and deaths were found. LIMITATIONS: Single-centre retrospective design, non-randomized sample, small sample size. CONCLUSION: Although elderly LSS patients (≥ 80 years old) are less fit and have more comorbidities, satisfactory outcomes can be achieved with PELD, comparable to those of LSS patients < 80 years old, and without increased complications.

New drug approvals for 2022: Synthesis and clinical applications.

The U.S. Food and Drug Administration has approved a total of 37 new drugs in 2022, which are composed of 20 chemical entities and 17 biologics. In particular, 20 chemical entities, including 17 small molecule drugs, 1 radiotherapy, and 2 diagnostic agents, provide privileged scaffolds, breakthrough clinical benefits, and a new mechanism of action for the discovery of more potent clinical candidates. The structure-based drug development with clear targets and fragment-based drug development with privileged scaffolds have always been the important modules in the field of drug discovery, which could easily bypass the patent protection and bring about improved biological activity. Therefore, we summarized the relevant valuable information about clinical application, mechanism of action, and chemical synthesis of 17 newly approved small molecule drugs in 2022. We hope this timely and comprehensive review could bring about creative and elegant inspiration on the synthetic methodologies and mechanism of action for the discovery of new drugs with novel chemical scaffolds and extended clinical indications.

Additive Manufacturing of Engineered Living Materials with Bio-augmented Mechanical Properties and Resistance to Degradation.

Engineered living materials (ELMs) combine living cells with polymeric matrices to yield unique materials with programmable functions. While the cellular platform and the polymer network determine the material properties and applications, there are still gaps in our ability to seamlessly integrate the biotic (cellular) and abiotic (polymer) components into singular material, then assemble them into devices and machines. Herein, we demonstrated the additive-manufacturing of ELMs wherein bioproduction of metabolites from the encapsulated cells enhanced the properties of the surrounding matrix. First, we developed aqueous resins comprising bovine serum albumin (BSA) and poly(ethylene glycol diacrylate) (PEGDA) with engineered microbes for vat photopolymerization to create objects with a wide array of 3D form factors. The BSA-PEGDA matrix afforded hydrogels that were mechanically stiff and tough for use in load-bearing applications. Second, we demonstrated the continuous in situ production of L-DOPA, naringenin, and betaxanthins from the engineered cells encapsulated within the BSA-PEGDA matrix. These microbial metabolites bioaugmented the properties of the BSA-PEGDA matrix by enhancing the stiffness (L-DOPA) or resistance to enzymatic degradation (betaxanthin). Finally, we demonstrated the assembly of the 3D printed ELM components into mechanically functional bolts and gears to showcase the potential to create functional ELMs for synthetic living machines.

Effects of metformin on Streptococcus suis LuxS/AI-2 quorum sensing system and biofilm formation.

Streptococcus suis (S. suis) regulates biofilm formation through LuxS/AI-2 quorum sensing system, increasing drug resistance and exacerbating infection. The anti-hyperglycaemic agent metformin has anti-bacterial and anti-biofilm activities. This study aimed to investigate the anti-biofilm and anti-quorum sensing activity of metformin in S. suis. We first determined the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of metformin on S. suis. The results indicated that metformin showed no obvious inhibitory or bactericidal effect. Crystal violet staining showed that metformin significantly inhibited the formation of S. suis biofilm at sub-MIC concentration, which was also confirmed by scanning electron microscopy. Then, we quantified the AI-2 signal molecules in S. suis, and the results showed that metformin had a significant inhibitory effect on the production of AI-2 signal in S. suis. Inhibition of enzyme activity and molecular docking experiments showed that metformin has a significant binding activity to LuxS protein. In addition, qRT-PCR results showed that metformin significantly down-regulated the expression of AI-2 synthesis-related genes luxS and pfs, and adhesion-related genes luxS, pfs, gapdh, sly, fbps, and ef. Western blotting also showed that metformin significantly reduced the expression of LuxS protein. Our study suggests that metformin seems to be a suitable candidate for the inhibition of S. suis LuxS/AI-2 QS system and prevention of biofilm formation, which provided a new idea for the prevention and control of S. suis.

Enhancing long-term storage and stability of engineered living materials through desiccant storage and trehalose treatment.

Engineered living materials (ELMs) have broad applications for enabling on-demand bioproduction of compounds ranging from small molecules to large proteins. However, most formulations and reports lack the capacity for storage beyond a few months. In this study, we develop an optimized procedure to maximize stress resilience of yeast-laden ELMs through the use of desiccant storage and 10% trehalose incubation before lyophilization. This approach led to over 1-year room temperature storage stability across a range of strain genotypes. In particular, we highlight the superiority of exogenously added trehalose over endogenous, engineered production in yielding robust preservation resilience that is independent of cell state. This simple, effective protocol enables sufficient accumulation of intracellular trehalose over a short period of contact time across a range of strain backgrounds without requiring the overexpression of a trehalose importer. A variety of microscopic analysis including µ-CT and confocal microscopy indicate that cells form spherical colonies within F127-BUM ELMs that have variable viability upon storage. The robustness of the overall procedure developed here highlights the potential for widespread deployment to enable on-demand, cold-chain independent bioproduction.

Direct Alkylation of Quinoxalinones with Boracene-Based Alkylborate under Visible Light Irradiation.

An efficient synthesis of a variety of 3-alkyl quinoxalinones via C-H direct alkylation by photoredox catalysis between quinoxalinones and alkylborates is reported. A range of quinoxalinones was tolerated well. This visible-light photocatalysis reaction allows access to structurally diverse 3-alkyl quinoxalinones in good to excellent yields. The practicality of this protocol was demonstrated by the concise synthesis of a potential bioactive nonpeptide angiotensin II receptor antagonist.

Targeting histone deacetylases for heart diseases.

Histone deacetylases (HDACs) are responsible for the deacetylation of lysine residues in histone or non-histone substrates, leading to the regulation of many biological functions, such as gene transcription, translation and remodeling chromatin. Targeting HDACs for drug development is a promising way for human diseases, including cancers and heart diseases. In particular, numerous HDAC inhibitors have revealed potential clinical value for the treatment of cardiac diseases in recent years. In this review, we systematically summarize the therapeutic roles of HDAC inhibitors with different chemotypes on heart diseases. Additionally, we discuss the opportunities and challenges in developing HDAC inhibitors for the treatment of cardiac diseases.

Recent development of multi-target VEGFR-2 inhibitors for the cancer therapy.

Vascular epidermal growth factor receptor-2 (VEGFR-2), as an important tyrosine transmembrane protein, plays an important role in regulating endothelial cell proliferation and migration, regulating angiogenesis and other biological functions. VEGFR-2 is aberrantly expressed in many malignant tumors, and it is also related to the occurrence, development, and growth of tumors and drug resistance. Currently, there are nine VEGFR-2 targeted inhibitors approved by US.FDA for clinical use as anticancer drugs. Due to the limited clinical efficacy and potential toxicity of VEGFR inhibitors, it is necessary to develop new strategies to improve the clinical efficacy of VEGFR inhibitors. The development of multitarget therapy, especially dual-target therapy, has become a hot research field of cancer therapy, which may provide an effective strategy with higher therapeutic efficacy, pharmacokinetic advantages and low toxicity. Many groups have reported that the therapeutic effects could be improved by simultaneously inhibiting VEGFR-2 and other targets, such as EGFR, c-Met, BRAF, HDAC, etc. Therefore, VEGFR-2 inhibitors with multi-targeting capabilities have been considered to be promising and effective anticancer agents for cancer therapy. In this work, we reviewed the structure and biological functions of VEGFR-2, and summarized the drug discovery strategies, and inhibitory activities of VEGFR-2 inhibitors with multi-targeting capabilities reported in recent years. This work might provide the reference for the development of VEGFR-2 inhibitors with multi-targeting capabilities as novel anticancer agents.

Comparative efficacy and complications of single and dual growing rods for early-onset scoliosis: an updated meta-analysis.

PURPOSE: This updated meta-analysis aimed to compare single and dual growing rods, including both traditional growing rod and magnetically controlled growing rod (MCGR) used in the treatment of early-onset scoliosis (EOS) with regard to deformity correction, spinal growth, and complications. METHODS: This meta-analysis was performed according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines using articles extracted from PubMed, EMBASE databases, and Cochrane Library databases. Only articles reporting the complications and the imaging parameters before and after growing rods in the patients diagnosed with EOS were included. We extracted and statistically analyzed the data deemed relevant for this study, and used the Newcastle-Ottawa Scale to assess the risk of bias in each study. Data synthesis and statistical analyses were performed using R software. RESULTS: Fifteen eligible articles containing 409 participants (n = 185, single growing rods; n = 224, dual growing rods) were identified. The meta-analysis found no significant differences in the preoperative and postoperative major Cobb angle, T1-S1 distance, thoracic kyphosis, and coronal balance between single and dual rods groups. The final follow-up major Cobb angle (P = 0.01; standardized mean difference, - 0.42 [95% confidence interval (CI), - 0.74 to - 0.10]; I2 = 23%) was significantly smaller in dual rods group than single-rod group. However, no significant differences in the correction rate of angle (major Cobb angle and kyphosis angle) and changes in the T1-S1 distance between the two groups were observed. Moreover, there were no significant differences in the metalwork failure, infection, or proximal junctional kyphosis between single and dual rods groups. However, total complications (P = 0.03; risk ratio (RR), 0.79 [95% CI, 0.63-0.98]; I2 = 29%) and distraction failure in MCGR (P = 0.04; RR, 0.38 [95% CI, 0.14-0.98]; I2 = 11%) were significantly lower in dual rods group than single-rod group. CONCLUSION: This updated meta-analysis found that patients with dual growing rods had fewer complications, especially distraction failure in MCGR, than those with single growing rod. However, none of deformity correction, spinal growth, or other complications differed between single and dual growing rods. Therefore, we believe that dual growing rods do not provide strong advantages over single growing rod in the treatment of EOS.

Morphometric change in intervertebral foramen after percutaneous endoscopic lumbar foraminotomy: an in vivo radiographic study based on three-dimensional foramen reconstruction.

PURPOSE: This study aimed to perform in vivo three-dimensional (3D) quantitative measurements of morphometric changes in the foramen in patients with lumbar foraminal stenosis (LFS) undergoing percutaneous endoscopic lumbar foraminotomy (PELF) and investigate the relationship between anatomical changes in the foramen and clinical outcomes. METHODS: We retrospectively reviewed consecutive patients with LFS treated with PELF between January 2016 and September 2020 at our centre. Clinical outcomes were evaluated. Foraminal volume (FV) and foraminal minimal area (FMA) were calculated using a novel vertebral and foramen segmentation method. A comparison of the anatomical parameters of the foramen were conducted between the satisfied and unsatisfied groups divided based on the modified MacNab criteria. RESULTS: A total of 26 eligible patients with a mean follow-up of 3.6 years were enrolled. A significant increase was found in overall FV (71.5%) from 1.436 ± 0.396 to 2.464 ± 0.719 cm3 (P < 0.001) and FMA (109.5%) from 0.849 ± 0.207 to 1.780 ± 0.524 cm2. All clinical outcomes were significantly improved (P < 0.001) after PELF. No significant difference was found in changes in neither FV nor FMA between the two groups. CONCLUSION: Clinical results and foraminal dimensions improved significantly after PELF, indicating that PELF was a prominent technique suitable for LFS because of the direct decompression at impingement structures. No relationship was found between morphometric changes and clinical outcomes, revealing that full-scale endoscopic decompression is necessary and adequate for LFS, and unsatisfactory outcomes are less likely to result from decompression procedure.

Synthesis, In Vitro, and In Vivo Investigations of Pterostilbene-Tethered Analogues as Anti-Breast Cancer Candidates.

Pterostilbene has been found to be an active scaffold with anti-breast cancer (BC) action. In this study, fourteen pterostilbene-tethered analogues (2A-2N) were prepared and screened in vitro against MDA-MB-231 and MCF-7 cells. Meanwhile, their structures were characterized using 1H-NMR, 13C-NMR, and HRMS (ESI) spectroscopy techniques. Among them, analogue 2L displayed the most potent anti-proliferation effect on MDA-MB-231 (IC50 = 10.39 µM) and MCF-7 cells (IC50 = 11.73 µM). Furthermore, the meaningful structure-activity relationships suggested that the introduction of a saturated six-membered nitrogen heterocyclic ring into the side chain favored anti-BC capacity. Biological observations indicated that 2L could cause the typical morphological changes in apoptosis, namely an increase in reactive oxygen species level and a loss of mitochondrial membrane potential in BC cells. Importantly, 2L could induce mitochondrial-mediated apoptosis by regulating the expression of caspase-related proteins. Consistent with the results of our in vitro study, 2L apparently inhibited tumor growth in MDA-MB-231 xenograft mice without obvious toxicity. These findings revealed that 2L is expected to be a promising anti-BC lead compound that merits further investigations.

A Review of the Regulatory Mechanisms of N-Myc on Cell Cycle.

Neuroblastoma has obvious heterogeneity. It is one of the few undifferentiated malignant tumors that can spontaneously degenerate into completely benign tumors. However, for its high-risk type, even with various intensive treatment options, the prognosis is still unsatisfactory. At the same time, a large number of research data show that the abnormal amplification and high-level expression of the MYCN gene are positively correlated with the malignant progression, poor prognosis, and mortality of neuroblastoma. In this context, this article explores the role of the N-Myc, MYCN gene expression product on its target genes related to the cell cycle and reveals its regulatory network in promoting tumor proliferation and malignant progression. We hope it can provide ideas and direction for the research and development of drugs targeting N-Myc and its downstream target genes.

Exploring the Mechanism of Hepatotoxicity Induced by Dictamnus dasycarpus Based on Network Pharmacology, Molecular Docking and Experimental Pharmacology.

The root bark of Dictamnus dasycarpus Turcz is a traditional Chinese medicine, Dictamni Cortex (DC), which is mainly used in the clinical treatment of skin inflammation, eczema, rubella, rheumatism, and gynecological inflammation. Unexpectedly, there are some cases of liver injury after the administration of DC. However, the mechanism of hepatotoxicity remains ambiguous. The aim of this study was to explore the mechanism and substance bases of DC hepatotoxicity based on network pharmacology and molecular docking, verified through pharmacological experiments. Partial prototype components and metabolites in vivo of quinoline alkaloids from DC were selected as candidate compounds, whose targets were collected from databases. Network pharmacology was applied to study the potential hepatotoxic mechanism after correlating the targets of candidate compounds with the targets of hepatotoxicity. Molecular docking was simulated to uncover the molecular mechanism. Furthermore, the hepatotoxicity of the extract and its constituents from DC was evaluated in vivo and in vitro. We constructed the "potential toxic components-toxic target-toxic pathway" network. Our results showed that the targets of DC included CYP1A2 and GSR, participating in heterologous steroid metabolism, REDOX metabolism, drug metabolism, heterocyclic metabolic processes, the synthesis of steroid hormone, cytochrome P450 metabolism, chemical carcinogens and bile secretion pathways. In vitro and in vivo experiments displayed that DC could result in a decrease in GSH-Px and oxidative stress, simultaneously inhibiting the expression of CYP1A2 and inducing hepatotoxicity. These results further indicated the mechanism of hepatotoxicity induced by Dictamnus dasycarpus, providing a basic theory to explore and prevent hepatotoxicity in the clinical usage of Dictamnus dasycarpus.

A single amino acid mutation in the mouse MEIG1 protein disrupts a cargo transport system necessary for sperm formation.

Mammalian spermatogenesis is a highly coordinated process that requires cooperation between specific proteins to coordinate diverse biological functions. For example, mouse Parkin coregulated gene (PACRG) recruits meiosis-expressed gene 1 (MEIG1) to the manchette during normal spermiogenesis. Here we mutated Y68 of MEIG1 using the CRISPR/cas9 system and examined the biological and physiological consequences in mice. All homozygous mutant males examined were completely infertile, and sperm count was dramatically reduced. The few developed sperm were immotile and displayed multiple abnormalities. Histological staining showed impaired spermiogenesis in these mutant mice. Immunofluorescent staining further revealed that this mutant MEIG1 was still present in the cell body of spermatocytes, but also that more MEIG1 accumulated in the acrosome region of round spermatids. The mutant MEIG1 and a cargo protein of the MEIG1/PACRG complex, sperm-associated antigen 16L (SPAG16L), were no longer found to be present in the manchette; however, localization of the PACRG component was not changed in the mutants. These findings demonstrate that Y68 of MEIG1 is a key amino acid required for PACRG to recruit MEIG1 to the manchette to transport cargo proteins during sperm flagella formation. Given that MEIG1 and PACRG are conserved in humans, small molecules that block MEIG1/PACRG interaction are likely ideal targets for the development of male contraconception drugs.