A durable 4-1BB-based CD19 CAR-T cell for treatment of relapsed or refractory non-Hodgkin lymphoma

Objective:Previous studies reported that 4-1BB-based CD19 chimeric antigen receptor(CAR)-T cells were more beneficial for the clinical outcomes than CD28-based CAR-T cells,especially the lower incidence rate of severe adverse events.However,the median progression-free survival(mPFS)of 4-1BB-based product Kymriah was shorter than that of CD28-based Yescarta(2.9 months vs.5.9 months),suggesting that Kymriah was limited in the long-term efficacy.Thus,a safe and durable 4-1BB-based CD19 CAR-T needs to be developed.Methods:We designed a CD19-targeted CAR-T(named as IM19)which consisted of an FMC63 scFv,4-1BB and CD3ζintracellular domain and was manufactured into a memory T-enriched formulation.A phase I/II clinical trial was launched to evaluate the clinical outcomes of IM19 in relapsed or refractory(r/r)B cell non-Hodgkin lymphoma(B-NHL).Dose-escalation investigation(at a dose of 5×10^(5)/kg,1×10^(6)/kg and 3×106/kg)was performed in 22 r/r B-NHL patients.All patients received a single infusion of IM19 after 3-day conditional regimen.Results:At month 3,the overall response rate(ORR)was 59.1%,the complete response rate(CRR)was 50.0%.The mPFS was 6 months and the 1-year overall survival rate was 77.8%.Cytokine release syndrome(CRS)occurred in 13 patients(59.1%),with 54.5%of grade 1−2 CRS.Only one patient(4.5%)experienced grade 3 CRS and grade 3 neurotoxicity.Conclusions:These results demonstrated the safety and durable efficacy of a 4-1BB-based CD19 CAR-T,IM19,which is promising for further development and clinical investigation.

Sensing Traction Strain Induces Cell-Cell Distant Communications

Mechanobiology has been a highly recognized field in studying the importance of physical forces in physiologies at the molecular,cellular,tissue,organ and body-levels.Beside the intensive work focusing on the fine local biomechanical forces,the long-range force which can propagate through a relatively distant scale(in hundreds of micrometers and beyond)has been an intriguing topic with increasing attentions in recent years.The collective functions at cell population level often rely on cell-cell communications with or without direct contacts.Recent progresses including our own work indicate that the long-range biomechanical force propagating across scales far beyond single cell size may reserve the capability to trigger coordinative biological responses within cell population.Whether and how cells communicate mechanically in a distant manner remains largely to be explored.In respiratory system,the mechanical property of airway smooth muscle(ASM)is associated with asthma attack with prolonged contraction during airway hyper-responsiveness.In this work,we found that ASM cells rapidly self-assembled into a well-constructed network on 3D matrigel containing type I collagen(COL I),which required the collective functions and coordination of thousands of cells completed within 12-16 hours.Cells were assembled with aligned actin stress fibers and elongated nuclei.The assembling process relied on the long-range mechanical forces across the matrix to direct cell-cell distant interactions.We further found that single ASM cells could rapidly initiate multiple buds precisely pointing to neighboring cells in distance,which relied on cell traction force and force strain on the matrix.Beads tracking assay demonstrated the long-range transmission of cellular traction force to distant locations,and modeling of maximum strain distribution on matrix by finite element method predicted the consistency with cell directional protrusions and movements in experiments.Cells could sense each other in distance to move directionally on both non-fibrous matrigel and in much more efficient way when containing COL I.Cells recruited COL I from the hydrogel to build nearly identical COL I fibrous network to mechanically stabilize the cell network.Our results revealed that ASM cells can sense the traction strain transmitted through matrix to initiate distant communications and rapidly coordinate the network assembly at the population level through active cell-matrix interactions.As an interesting phenomenon,cells sound able to’make phone call’via the role of long-range mechanical force.In summary,this work demonstrated that long-range biomechanical force facilitates the collective functions of ASM cell population for network assembly.The cells reacted to traction strain on the matrix for distant communications,which resulted in directional budding and movement.Fibrous COL I had important roles in facilitating the efficiency of force transmission to induce the assembly and stabilizing the cell network.This work has helped advance the understanding of the feature andfunction of long-range biomechanical force at the cell population level.The observed high mechano-sensitivity of ASM cells might suggest a re-enforced feedback of enhanced contraction by excessive ASM under asthmatic condition.

Mechanics of water pore formation in lipid membrane under electric field

Transmembrane water pores are crucial for substance transport through cell membranes via membrane fusion, such as in neural communication. However, the molecular mechanism of water pore formation is not clear. In this study, we apply all-atom molecular dynamics and bias-exchange metadynamics simulations to study the process of water pore formation under an electric field. We show that water molecules can enter a membrane under an electric field and form a water pore of a few nanometers in diameter. These water molecules disturb the interactions between lipid head groups and the ordered arrangement of lipids. Following the movement of water molecules, the lipid head groups are rotated and driven into the hydrophobic region of the membrane. The reorientated lipid head groups inside the membrane form a hydrophilic surface of the water pore. This study reveals the atomic details of how an electric field influences the movement of water molecules and lipid head groups, resulting in water pore formation.

Effect of antisense oligonucleotide targeting bFGF on apoptosis of hepatoma cells

Objective： To investigate the cell cycle changes of hepatoma cells and the role of antisense oligonucleotide targeting bFGF. Methods： Inhibition of bFGF protein expression was investigated by confocal microscopy analysis and Western blot in the best condition of transfecting antisense oligonucleotide targeting bFGF. Cell cycle and apoptosis were detected with flow cytometry analysis. Results： Treatment with antisense oligonucleotide of bFGF not only reduced the expression of bFGF by confocal microscopy and Western blot analysises, but also increased the apoptosis of HepG2 cells （ P 〈 0.01）. Conclusion： bFGF may take part in apoptosis regulation of hepatoma cells and be used as a target of hepatocellular carcinoma therapy.

Cell Mechanics Regulates the Dynamic Anisotropic Remodeling of Fibril Matrix at Large Scale

Living tissues often have anisotropic and heterogeneous organizations, in which developmental processes are coordinated by cells and extracellular matrix modeling. Cells have the capability of modeling matrix in long distance;however, the biophysical mechanism is largely unknown. We investigated the dynamic remodeling of collagen I (COL) fibril matrix by cell contraction with designed patterns of cell clusters. By considering cell dynamic contractions, our molecular dynamics simulations predicted the anisotropic patterns of the observed COL bundling in experiments with various geometrical patterns without spatial limitation. The pattern of COL bundling was closely related to the dynamic remodeling of fibril under cell active contraction. We showed that cell cytoskeletal integrity (actin filaments and microtubules), actomyosin contractions, and endoplasmic reticulum calcium channels acting as force generations and transductions were essential for fiber bundling inductions, and membrane mechanosensory components integrin and Piezo played critical roles as well. This study revealed the underlying mechanisms of the cell mechanics-induced matrix remodeling in large scales and the associated cellular mechanism and should provide important guidelines for tissue engineering in potential biomedical applications.

Single-vesicle imaging quantifies calcium’s regulation of nanoscale vesicle clustering mediated by α-synuclein

Although numerous studies have shown that the proteinα-synuclein(α-Syn)plays a central role in Parkinson’s disease,dementia with Lewy bodies,and other neurodegenerative diseases,the protein’s physiological function remains poorly understood.Furthermore,despite recent reports suggesting that,under the influence of Ca^(2+),α-Syn can interact with synaptic vesicles,the mechanisms underlying that interaction are far from clear.Thus,we used single-vesicle imaging to quantify the extent to which Ca^(2+)regulates nanoscale vesicle clustering mediated by α-Syn.Our results revealed not only that vesicle clustering requiredα-Syn to bind to anionic lipid vesicles,but also that different concentrations of Ca^(2+)exerted different effects on howα-Syn induced vesicle clustering.In particular,low concentrations of Ca^(2+)inhibited vesicle clustering by blocking the electrostatic interaction between the lipid membrane and the N terminus of α-Syn,whereas high concentrations promoted vesicle clustering,possibly due to the electrostatic interaction between Ca^(2+)and the negatively charged lipids that is independent of α-Syn.Taken together,our results provide critical insights intoα-Syn’s physiological function,and how Ca^(2+) regulates vesicle clustering mediated by α-Syn.