BRAF and KRAS mutations in metastatic colorectal cancer:future perspectives for personalized therapy

Colorectal cancer(CRC)is one of the most commonly diagnosed cancers worldwide and 30%of patients with CRC experience metastasis.Patients with metastatic colorectal cancer(mCRC)have a 5-year overall survival rate of<10%.V-raf murine sarcoma viral oncogene homolog B1(BRAF)and V-Ki-ras2 Kirsten ratsarcoma viral oncogene homolog(KRAS)mutations are mostly studied in mCRC,as clinical trials found that first-line chemotherapy with anti-epidermal growth factor receptor agent confers limited efficacy for mCRC.Treatment decisions for early-stage mCRC do not consider BRAF or KRAS mutations,given the dramatically poor prognosis conferred by these mutations in clinical trials.Thus,it is necessary to identify patients with mCRC harboring BRAF or KRAS mutations to formulate rational therapeutic strategies to improve prognosis and survival.BRAF and KRAS mutations occur in10%and44%of patients with mCRC,respectively.Although the survival rate of patients with mCRC has improved in recent years,the response and prognosis of patients with the aforementioned mutations are still poor.There is a substantial unmet need for prospective personalized therapies for patients with BRAF-or KRAS-mutant mCRC.In this review,we focus on BRAF and KRAS mutations to understand the mechanisms underlying resistance and improving the response rate,outcomes,and prognosis of patients with mCRC bearing these mutations and to discuss prospective personalized therapies for BRAF-and KRAS-mutant mCRC.

Biomarkers for diabetes prediction, diagnosis and personalized therapy

Diabetes mellitus （DM） is one of the most common ＇metabolic disorders in the world~ in which more than90% are grouped to type 2 DM （T2DM）.1 T2DM is characterized by decreased insulin sensitivity and impaired insulin secretion2 leading to hyperglycemia, and the serum glucose has been used as a golden standard for diabetes diagnosis. However, T2DM is a kind of disease involving defects of multiple organs, which cannot be distinguished through the measurement of the serum-glucose level. In addition, T2DM is a multiple-stage disease, which usually covers several decades from impaired plasma glucose to various complications. The serum-glucose level only reflects the consequence of multiole physiological disorders in the Riven stare.

Gene-modified leucoconcentrate for personalized ex vivo gene therapy in a mini pig model of moderate spinal cord injury

We previously demonstrated that gene-modified umbilical cord blood mononuclear cells overexpressing a combination of recombinant neurotrophic factors are a promising therapeutic approach for cell-mediated gene therapy for neurodegenerative diseases,neurotrauma,and stroke.In this study,using a mini pig model of spinal cord injury,we proposed for the first time the use of gene-modified leucoconcentrate prepared from peripheral blood in the plastic blood bag for personalized ex vivo gene therapy.Leucoconcentrate obtained from mini pig peripheral blood was transduced with a chimeric adenoviral vector(Ad5/35 F)that carried an enhanced green fluorescent protein(EGFP)reporter gene in the plastic blood bag.The day after blood donation,the mini pigs were subjected to moderate SCI and four hours post-surgery they were intravenously autoinfused with gene-modified leucoconcentrate.A week after gene-modified leucoconcentrate therapy,fluorescent microscopy revealed EGFP-expressing leucocytes in spinal cord at the site of contusion injury.In the spleen the groups of EGFP-positive cells located in the lymphoid follicles were observed.In vitro flow cytometry and fluorescent microscopy studies of the gene-modified leucoconcentrate samples also confirmed the production of EGFP by leucocytes.Thus,the efficacy of leucocytes transduction in the plastic blood bag and their migratory potential suggest their use for temporary production of recombinant biologically active molecules to correct certain pathological conditions.This paper presents a proof-of-concept of simple,safe and effective approach for personalized ex vivo gene therapy based on gene-modified leucoconcentrate autoinfusion.The animal protocols were approved by the Kazan State Medical University Animal Care and Use Committee(approval No.5)on May 27,2014.

Advances and future directions in keloid research:Pathogenesis,diagnosis and personalized treatment strategies

Keloids,which are abnormal manifestations of wound healing,can result in significant functional impairment and aesthetic deformities.The pathogenesis of keloids is multifaceted and complex and influenced by various factors,such as genetics,the environment,and immune responses.The evolution of keloid treatment has progressed from traditional surgical excision to a contemporary combination of therapies including injection and radiation treatments,among others.This article provides a comprehensive review of keloid pathogenesis and treatment,emphasizing the latest advances in the field.Ultimately,this review underscores the necessity for continued research to enhance our understanding of keloid pathogenesis and to devise more effective treatments for this challenging condition.

Changing the paradigm：the potential for targeted therapy in laryngeal squamous cell carcinoma

Laryngeal squamous cell carcinoma(LSCC) remains a highly morbid and fatal disease. Historically, it has been a model example for organ preservation and treatment stratification paradigms. Unfortunately, survival for LSCC has stagnated over the past few decades. As the era of next-generation sequencing and personalized treatment for cancer approaches, LSCC may be an ideal disease for consideration of further treatment stratification and personalization. Here, we will discuss the important history of LSCC as a model system for organ preservation, unique and potentially targetable genetic signatures of LSCC, and methods for bringing stratified, personalized treatment strategies to the 21^(st) century.

Therapeutic options for the management of pancreatic cancer

Since its initial characterization, pancreatic ductal adenocarcinoma has remained one of the most devastating and difficult cancers to treat. Pancreatic cancer is the fourth leading cause of death in the United States, resulting in an estimated 38460 deaths annually. With few screening tools available to detect this disease at an early stage, 94% of patients will die within five years of diagnosis. Despite decades of research that have led to a better understanding of the molecular and cellular signaling pathways in pancreatic cancer cells, few effective therapies have been developed to target these pathways. Other treatment options have included more sophisticated pancreatic cancer surgeries and combination therapies. While outcomes have improved modestly for these patients, more effective treatments are desperately needed. One of the greatest challenges in the future of treating this malignancy will be to develop therapies that target the tumor microenvironment and surrounding pancreatic cancer stem cells in addition to pancreatic cancer cells. Recent advances in targeting pancreatic stellate cells and the stroma have encouraged researchers to shift their focus to the role of desmoplasia in pancreatic cancer pathobiology in the hopes of developing newer-generation therapies. By combining novel agents with current cytotoxic chemotherapies and radiation therapy and personalizing them to each patient based on specific biomarkers, the goal of prolonging a patient&#x02019;s life could be achieved. Here we review the most effective therapies that have been used for the treatment of pancreatic cancer and discuss the future potential of therapeutic options.

Emerging molecular classifications and therapeutic implications for gastric cancer

Gastric cancer(GC) is a highly aggressive and life-threatening malignancy.Even with radical surgical removal and front-line chemotherapy,more than half of GCs locally relapse and metastasize at a distant site.The dismal outcomes reflect the ineffectiveness of a one-size fits-all approach for a highly heterogeneous disease with diverse etiological causes and complex molecular underpinnings.The recent comprehensive genomic and molecular profiling has led to our deepened understanding of GC.The emerging molecular classification schemes based on the genetic,epigenetic,and molecular signatures are providing great promise for the development of more effective therapeutic strategies in a more personalized and precise manner.To this end,the Cancer Genome Atlas(TCGA) research network conducted a comprehensive molecular evaluation of primary GCs and proposed a new molecular classification dividing GCs into four subtypes:Epstein-Barr virus-associated tumors,microsatellite unstable tumors,genomically stable tumors,and tumors with chromosomal instability.This review primarily focuses on the TCGA molecular classification of GCs and discusses the implications on novel targeted therapy strategies.We believe that these fundamental findings will support the future application of targeted therapies and will guide our efforts to develop more efficacious drugs to treat human GCs.

Culture-guided treatment approach for Helicobacter pylori infection:Review of the literature

The progressive loss of efficacy of standard eradication therapies has made the treatment of Helicobacter pylori (H. pylori) more challenging than ever. Endoscopic-guided antibiotic susceptibility testing had previously been suggested to guide treatment after failure of second-line therapies. However, its role has expanded over the years, in accordance with the current Maastricht Guidelines. Several authors have dealt with this topic, developing both efficacy trials and cost-effectiveness trials against resistant H. pylori infections as well as infections in na&#x000ef;ve patients. However, results are not homogeneous enough to provide definite advice, because antibiotic resistance is not the only reason for treatment failure. Moreover, the culture-guided approach is surrounded by many practical issues, such as the availability of both endoscopy units and microbiology laboratories, and the need for a standard of quality that cannot be satisfied everywhere. Finally, pre-treatment susceptibility testing should be part - and not the only weapon - of a targeted, personalized strategy to overcome H. pylori infection.

The study on psychological resilience of tinnitus and associated influencing factors

The association between tinnitus and psychological resilience is an underdeveloped area of research.This cross-sectional study investigated such associations and factors potentially affecting resilience in 61 patients.Demographic and psychometric data were collected by questionnaires.The ConnoreDavidson Resilience Scale(CD-RISC),Medical Coping Modes Questionnaire(MCMQ),Satisfaction with Life Scale(SWLS),General Self-Efficacy Scale(GSES),Big Five Inventory(BFI)and Perceived Social Support Scale(PSSS)were completed by participants.Data were analyzed using independent t-test and Pearson's correlation analysis and multiple linear regression modeling.The CD-RISC score was relatively low(66.97±15.71),negatively correlated with tinnitus(r=0.276,p<0.001)and associated with age(r=0.270,P<0.001).As protective factors,SWLS(r=0.486,p<0.001),GSES(r=0.555,p<0.001),PSSS(r=0.538,p<0.001)and extraversion were positively correlated with CD-RISC and BFI scores(r=0.287,p<0.001).We also detected a negative correlation with neuroticism(r=0.395,p<0.001),which is a known risk factor for worse CD-RISC scores.Identifying protective and risk factors for psychological resilience can be used to predict treatment outcomes in tinnitus patients,which will help devise personalized solutions and improve patients'quality of life.

Stimulation and Control of Homeostasis

Healthy homeostasis is a principal driving force of the dynamic equilibrium of living organisms. The dynamical basis of homeostasis is the complex and interconnected feedback mechanisms, which are fundamentally governed by the nervous system, mainly the balance of the sympathetic and parasympathetic controlling actions. The balancing regulation is well presented in the heart’s sinus node and can be measured by the time-domain heart-rate variation (HRV) of its frequency domain to analyze the constitutional frequencies of the variation. This last is a fluctuation that shows 1/f time fractal arrangement (f is the composing frequency). The time-fractal arrangement could depend on the structural fractal of the His-Purkinje system of the heart and personally modify the HRV. The cancers gradually destroy the homeostatic harmony, starting locally and finishing systemically. The controlling activity of vagus-nerve changes the HRV or the power density spectrum of the signal fluctuations in malignant development, presenting an appropriate control of the cancerous processes. The modified spectrum by a non-invasive radiofrequency treatment could arrest the tumor growth. An appropriate modulation could support the homeostatic control and force reconstructing of the broken complexity.

Induced pluripotent stem cells as an innovative model to study drug induced pancreatitis

Drug-induced pancreatitis is a gastrointestinal adverse effect concerning about 2%of drugs.The majority of cases are mild to moderate but severe episodes can also occur,leading to hospitalization or even death.Unfortunately,the mechanisms of this adverse reaction are still not clear,hindering its prevention,and the majority of data available of this potentially life-threatening adverse effect are limited to case reports leading to a probable underestimation of this event.In particular,in this editorial,special attention is given to thiopurine-induced pancreatitis(TIP),an idiosyncratic adverse reaction affecting around 5%of inflammatory bowel disease(IBD)patients taking thiopurines as immunosuppressants,with a higher incidence in the pediatric population.Validated biomarkers are not available to assist clinicians in the prevention of TIP,also because of the inaccessibility of the pancreatic tissue,which limits the possibility to perform dedicated cellular and molecular studies.In this regard,induced pluripotent stem cells(iPSCs)and the exocrine pancreatic differentiated counterpart could be a great tool to investigate the cellular and molecular mechanisms underlying the development of this undesirable event.This particular type of stem cells is obtained by reprogramming adult cells,including fibroblasts and leukocytes,with a set of transcription factors known as the Yamanaka’s factors.Maintaining unaltered the donors’genetic heritage,iPSCs represent an innovative model to study the mechanisms of adverse drug reactions in individual patients’tissues not easily obtainable from human probands.Indeed,iPSCs can differentiate under adequate stimuli into almost any somatic lineage,opening a new world of opportunities for researchers.Several works are already available in the literature studying liver,central nervous system and cardiac cells derived from iPSCs and adverse drug effects.However,to our knowledge no studies have been performed on exocrine pancreas differentiated from iPSCs and drug-induced pancreatitis,so far.Hence,in this editorial we focus specifically on the description of the study of the mechanisms of TIP by using IBD patient-specific iPSCs and exocrine pancreatic differentiated cells as innovative in vitro models.

Inverted Simulations Demonstrating Strong Ecological Fallacies in Cohort Studies

We start with a description of the statistical inferential framework and the duality between observed data and the true state of nature that underlies it. We demonstrate here that the usual testing of dueling hypotheses and the acceptance of one and the rejection of the other is a framework which can often be faulty when such inferences are applied to individual subjects. This follows from noting that the statistical inferential framework is predominantly based on conclusions drawn for aggregates and noting that what is true in the aggregate frequently does not hold for individuals, an ecological fallacy. Such a fallacy is usually seen as problematic when each data record represents aggregate statistics for counties or districts and not data for individuals. Here we demonstrate strong ecological fallacies even when using subject data. Inverted simulations, of trials rightly sized to detect meaningful differences, yielding a statistically significant p-value of 0.000001 （1 in a million） and associated with clinically meaningful differences between a hypothetical new therapy and a standard therapy, had a proportion of instances of subjects with standard therapy effect better than new therapy effects close to 30%. A ―winner take all‖ choice between two hypotheses may not be supported by statistically significant differences based on stochastic data. We also argue the incorrectness across many individuals of other summaries such as correlations, density estimates, standard deviations and predictions based on machine learning models. Despite artifacts we support the use of prospective clinical trials and careful unbiased model building as necessary first steps. In health care, high touch personalized care based on patient level data will remain relevant even as we adopt more high tech data-intensive personalized therapeutic strategies based on aggregates.

Personalized treatment based on mini patient-derived xenografts and WES/RNA sequencing in a patient with metastatic duodenal adenocarcinoma

Background:Treatment guidelines for a variety of cancers have been increasingly used in clinical practice,and have resulted in major improvement in patient outcomes.However,recommended regimens(even first-line treat-ments)are clearly not ideal for every patients.In the present study,we used mini patient-derived xenograft(mini-PDX)and next-generation sequencing to develop personalized treatment in a patient with metastatic duodenal adenocarcinoma.Methods:Resected metachronous metastatic tumor tissues were implanted into SCID mice to determine the sensitivity to a variety of drug regimens.Mutation profiles were assessed using both DNA whole-exome sequencing(DNA-WES)and RNA sequencing.The results of the analyses were used to select optimal treatment for the patient with metastatic duodenal adenocarcinoma.Results:Assessment with mini-PDX models took only 7 days.The results showed high sensitivity to S-1 plus cis-platin,gemcitabine plus cisplatin and everolimus alone.The patient received gemcitabine plus cisplatin initially,but the treatment was terminated due to toxicity.The patient was then switched to treatment with S-1 alone.The overall disease-free survival was 34 months.DNA-WES and RNA sequencing identified KRAS mutation(A146T),TP53(C229Yfs*10)and RICTOR amplification in the metastatic duodenal adenocarcinoma.These findings provided further support to the results of the mini-PDX,and suggest mTOR inhibitors should be used if and when relapse eventually occurs in this patient.Conclusions:Mini-PDX model combined with WES/RNA sequencing can rapidly assess drug sensitivity in cancer patients and reveal key genetic alterations.Further research on this technology for personalized therapy in patients with refractory malignant tumors is warranted.

Guided chemotherapy based on patient-derived mini-xenograft models improves survival of gallbladder carcinoma patients

Background:Gallbladder carcinoma is highly aggressive and resistant to chemotherapy,with no consistent strategy to guide first line chemotherapy.However,patient-derived xenograft(PDX)model has been increasingly used as an effective model for in preclinical study of chemosensitivity.Methods:Mini-PDX model was established using freshly resected primary lesions from 12 patients with gallblad-der to examine the sensitivity with five of the most commonly used chemotherapeutic agents,namely gemcitabine,oxaliplatin,5-fluorouracil,nanoparticle albumin-bound(nab)-paclitaxel,and irinotecan.The results were used to guide the selection of chemotherapeutic agents for adjunctive treatment after the surgery.Kaplan-Meier method was used to compare overall survival(OS)and disease free survival(DFS)with 45 patients who received conventional chemo-therapy with gemcitabine and oxaliplatin.Results:Cell viability assays based on mini-PDX model revealed significant heterogeneities in drug responsiveness.Kaplan-Meier analysis showed that patients in the PDX-guided chemotherapy group had significantly longer median OS(18.6 months;95%CI 15.9-21.3 months)than patients in the conventional chemotherapy group(13.9 months;95%CI 11.7-16.2 months)(P=0.030;HR 3.18;95%CI 1.47-6.91).Patients in the PDX-guided chemotherapy group also had significantly longer median DFS(17.6 months;95%CI 14.5-20.6 months)than patients in the conventional chemotherapy group(12.0 months;95%CI 9.7-14.4 months)(P=0.014;HR 3.37;95%CI 1.67-6.79).Conclusion:The use of mini-PDX model to guide selection of chemotherapeutic regimens could improve the out-come in patients with gallbladder carcinoma.

Is nutritional status a new indicator to use in clinical practice for colorectal cancer patients?

In this editorial we comment on the interesting article by Liu et al.The topic of discussion is the need for a cost-effective and easy-to-use scoring system for predicting the prognosis of colorectal cancer patients.In this context,nutritional assessment plays a crucial role in the multimodal evaluation of patients.In particular,the controlling nutritional status score was found to be an effective tool in the clinical decision-making process,in order to customize treatment strategies and to improve patient outcomes.

Patient-derived tumor models and their distinctive applications in personalized drug therapy

Tumor models in vitro are conventional methods for developing anti-cancer drugs,evaluating drug delivery,or calculating drug efficacy.However,traditional cell line-derived tumor models are unable to capture the tumor heterogeneity in patients or mimic the interaction between tumors and their surroundings.Recently emerging patient-derived preclinical cancer models,including of patient-derived xenograft(PDX)model,circulating tumor cell(CTC)-derived model,and tumor organoids-on-chips,are promising in personalized drug therapy by reca-pitulating the complexities and personalities of tumors and surroundings.These patient-derived models have demonstrated potential advantages in satisfying the rigorous demands of specificity,accuracy,and efficiency necessary for personalized drug therapy.However,the selection of suitable models is depending on the specific therapeutic requirements dictated by cancer types,progressions,or the assay scale.As an example,PDX models show remarkable advantages to reconstruct solid tumors in vitro to understand drug delivery and metabolism.Similarly,CTC-derived models provide a sensitive platform for drug testing in advanced-stage patients,while also facilitating the development of drugs aimed at suppressing tumor metastasis.Meanwhile,the demand for large-scale testing has promoted the development of tumor organoids-on-chips,which serves as an optimal tool for high-throughput drug screening.This review summarizes the establishment and development of PDX,CTC-derived models,and tumor organoids-on-chips and addresses their distinctive advantages in drug discovery,sensitive testing,and screening,which demonstrate the potential to aid in the selection of suitable models for fundamental cancer research and clinical trials,and further developing the personalized drug therapy.

Dosage individualization in children:integration of pharmacometrics in clinical practice

Background:Children are in a continuous and dynamically changing state of growth and development.A thorough understanding of developmental pharmacokinetics(PK)and pharmacodynamics(PD)is required to optimize drug therapy in children.Data sources:Based on recent publications and the experience of our group,we present an outline on integrating pharmacometrics in pediatric clinical practice to develop evidence-based personalized pharmacotherapy.Results:Antibiotics in septic neonates and immunosuppressants in pediatric transplant recipients are provided as proof-of-concept to demonstrate the utility of pharmacometrics in clinical practice.Dosage individualization based on developmental PK-PD model has potential benefits of improving the efficacy and safety of drug therapy in children.Conclusion:The pharmacometric technique should be better developed and used in clinical practice to personalize drug therapy in children in order to decrease variability of drug exposure and associated risks of overdose or underdose.

Progress in the Application of Ovarian and Fallopian Tube Organoids

As an innovative in vitro culture model,organoids have been established by cell sorting and subsequent culture in three-dimensional culture systems.Organoids can be derived from induced pluripotent stem cells or organ-restricted adult stem cells.Compared with traditional two-dimensional cell culture models and patient-derived xenograft models,organoids possess long-term genetic stability and can better retain the characteristics of source tissues or organs.These advantages have led to the increased use of ovarian and fallopian tube organoids in various fields of research,including cell differentiation and development,establishment of disease occurrence and progression models,tissue regeneration and reconstruction,individual drug screening,immune cell co-culture,and maternal-fetal medicine.This review briefly summarizes the recent progress in the application of ovarian and fallopian tube organoids in the field of obstetrics and gynecology.

Circulating tumor cells and the metastatic process: the complexity of malignancy

Despite improvements achieved in terms of early detection and therapeutic approach,metastatic breast cancer remains one of the principal worldwide causes of death.In recent years,due to the heterogeneous response of each patient to chemotherapy,clinical research highlights the need of a personalized approach.Circulating tumor cells(CTCs)represents a promising tool for this purpose.Unfortunately,even if their correlation with sever-ity,outcome and metastatic nature of the tumor has been established,several issues,mainly concerning their characterization and isolation,need to be solved.In this review,latest knowledge on CTCs and metastatic pro-cess in breast cancer were analyzed,aiming to understand their clinical utility and validity for a prospective ther-apeutic scenario.

Characterization of drug responses of mini patient-derived xenografts in mice for predicting cancer patient clinical therapeutic response

Background:Patient-derived organoids and xenografts(PDXs)have emerged as powerful models in functional diag-nostics with high predictive power for anticancer drug response.However,limitations such as engraftment failure and time-consuming for establishing and expanding PDX models followed by testing drug efficacy,and inability to subject to systemic drug administration for ex vivo organoid culture hinder realistic and fast decision-making in selecting the right therapeutics in the clinic.The present study aimed to develop an advanced PDX model,namely MiniPDX,for rapidly testing drug efficacy to strengthen its value in personalized cancer treatment.Methods:We developed a rapid in vivo drug sensitivity assay,OncoVee®MiniPDX,for screening clinically relevant regimens for cancer.In this model,patient-derived tumor cells were arrayed within hollow fiber capsules,implanted subcutaneously into mice and cultured for 7 days.The cellular activity morphology and pharmacokinetics were systematically evaluated.MiniPDX performance(sensitivity,specificity,positive and negative predictive values)was examined using PDX as the reference.Drug responses were examined by tumor cell growth inhibition rate and tumor growth inhibition rate in PDX models and MiniPDX assays respectively.The results from MiniPDX were also used to evaluate its predictive power for clinical outcomes.Results:Morphological and histopathological features of tumor cells within the MiniPDX capsules matched those both in PDX models and in original tumors.Drug responses in the PDX tumor graft assays correlated well with those in the corresponding MiniPDX assays using 26 PDX models generated from patients,including 14 gastric cancer,10 lung cancer and 2 pancreatic cancer.The positive predictive value of MiniPDX was 92%,and the negative predictive value was 81%with a sensitivity of 80%and a specificity of 93%.Through expanding to clinical tumor samples,Min-iPDX assay showed potential of wide clinical application.Conclusions:Fast in vivo MiniPDX assay based on capsule implantation was developed-to assess drug responses of both PDX tumor grafts and clinical cancer specimens.The high correlation between drug responses of paired MiniPDX and PDX tumor graft assay,as well as translational data suggest that MiniPDX assay is an advanced tool for personalized cancer treatment.