Autosomal recessive primary microcephaly (MCPH) is defined by congenital microcephaly and associated mental retardation with head circumference of the affected individual at least 3 standard deviations below age- and sex-means. It is a disorder of abnormal fetal brain growth which is a consequence of impaired neurogenesis. It is genetically heterogeneous with seven known loci and genes for all the seven loci have been identified: MCPH-1-MCPH1, MCPH2-WDR62, MCPH3-CDK5RAP2, MCPH4-CEP152, MCPH5-ASPM, MCPH6-CENPJ, and MCPH7-STIL. All the seven MCPH proteins localize at the centrosome. Apart from MCPH, many other proteins associated with the phenotype microcephaly have been localized to the centrosome or linked to it functionally. For example, Microcephalic osteodysplastic primordial dwarfism type II protein PCNT and Seckel syndrome protein ATR are also centrosomal proteins. All of the above findings show the importance of centrosomal proteins as the key players in neurogenesis and brain development. However, the exact mechanism as to how the loss-of-function of these proteins leads to microcephaly remains to be elucidated. The study of MCPH genes can also provide insights into the basics of neurogenesis that lead to a normal brain size. The most common cause of MCPH is mutations in the ASPM (abnormal spindle-like, microcephaly-associated protein) gene. The main aim of this study was to gain insight into the function of ASPM using the yeast two-hybrid technique. The main findings of the study are listed below. To find novel interacting proteins for SPM, a GAL4 based yeast two-hybrid system was used. The 3,477 amino acid long ASPM was divided into eight different baits and each bait was individually used for screening a human fetal brain cDNA library cloned in the pACT2 vector. To generate baits, the different regions were amplified from human fetal brain cDNA and cloned in-frame with the GAL4-DNA binding domain in the pGBKT7 vector. Screening with a C-terminus ASPM bait (pGBKT7-CTR) identified Angelman syndrome protein ubiquitin protein ligase E3A (UBE3A) as an ASPM interactor. A region of UBE3A from amino acids 639-875 was found to interact with ASPM. The identification of UBE3A as an ASPM interacting partner was interesting as more than 80% of Angleman syndrome patients are reported to have microcephaly. Screening with the baits pGBKT7-1.4 kb ASPM and pGBKT7-2.1 kb ASPM harboring parts of IQ domain identified calmodulin as an ASPM interating partner. The full length calmodulin was found to interact with the IQ domain of ASPM. The interactions identified in the yeast two-hybrid assay were confirmed in vivo by co-immunoprecipitation studies. For this, a rabbit polyclonal anti-ASPM antibody was raised against the N-terminal region of ASPM (from amino acids 544-1059). The specificity of the antibody was tested by Western blot analysis and immunofluorescence microscopy. ASPM antibody recognized the 410 KDa fulllength ASPM protein in lysates from human fetal tissues and different cell lines. Immunofluorescence analysis in HEK293 cells with the antibody revealed centrosomal staining of ASPM throughout mitosis and midbody staining in cytokinesis, as reported previously. Using antibodies against ASPM and UBE3A and human fetal kidney lysate, ASPM and UBE3A interaction was confirmed in vivo by co-immunoprecipitation. The interaction between ASPM and calmodulin was confirmed similarly. The relevance of the interaction between ASPM and UBE3A was pursued further Like ASPM, UBE3A localized to the centrosome throughout mitotic progression. ASPM levels were found to be unaffected upon overexpression of UBE3A in HEK293 cells, indicating that ASPM is not degraded by a UBE3A-dependent proteasomal pathway or the degradation may be spatial-temporal control. Further, immunofluorescence analysis of UBE3A overexpressing HEK293 cells revealed that UBE3A does not affect either the ASPM localization or its protein level at the centrosome. Synchronization of HEK293 cells in different cell cycle phases revealed that UBE3A is a cell cycle dependent protein and its level peaks in mitosis To explore the functional role of UBE3A’s increased level in mitosis, UBE3A was depleted in HEK293 cells with a shRNA construct and stable clones were generated. HEK293- UBE3A shRNA knockdown cells were examined for normal mitotic progession and spindle defects. There was a 3.81- to 5.52-fold increase in the frequency of anaphase/telophase cells with missegregated chromosomes in UBE3A knockdown clones as compared to scrambled clones. Hence, we identified a definitive role of UBE3A in chromosome segregation. Defective chromosome segregation has been reported in many studies associated with microcephaly-related proteins. Interestingly, chromosome malfunctioning has also been reported in Drosophilia asp mutants (ASPM orthologue) and Celegans aspm-1 knockdown cells. Therefore, the loss of both ASPM and UBE3A leading to chromosome segregation defects reveals the existence of a molecular pathway common to both ASPM and UBE3A As a consequence of chromosome missegregation, UBE3A knockdown cells were found to undergo abnormal cytokinesis and apoptosis. The percentage of apoptotic cells in UBE3A knockdown clones was 1.25- to 3.04-fold higher as compared to scrambled clones. Interestingly, an extensive apoptosis has been found in the neural folds of MCPH7 gene STIL null mice embryos. Thus, the present study links Angleman syndrome protein UBE3A to ASPM, centrosome and mitosis for the first time.

No description available.

O gene PKHD1, mutado na doença renal policística autossômica recessiva, apresenta um padrão de splicing complexo associado a múltiplos transcritos alternativos. Neste trabalho estudamos o perfil de expressão de seu produto, poliductina. Análises por western blot revelaram produtos putativos de membrana de > 440 kDa e ~230 kDa, e de ~140 kDa em frações solúveis de rim, fígado e pâncreas. Estudos imunoistoquímicos mostraram marcação em ductos coletores renais e porção ascendente espessa da alça de Henle, em epitélios ductais biliar e pancreático e, no período embrionário, em broto ureteral, ductos biliar e pancreático e glândula salivar. Análises por imunofluorescência e microscopia imunoeletrônica sugerem que poliductina se localize em cílio apical primário, membrana apical e citoplasma de células do ducto coletor. Nossos resultados indicam que PKHD1 codifica isoformas de membrana e solúveis / PKHD1, the gene mutated in autosomal recessive polycystic kidney disease, presents a complex splicing pattern, associated with multiple alternative transcripts. In this work we have studied the expression profile of its product, polyductin. Western blot analysis revealed putative membrane products of > 440 kDa and 230 kDa, and of ~140 kDa in soluble fractions in kidney, liver and pancreas. Immunohistochemistry studies showed staining in renal collecting duct and thick ascending limb of Henle, in biliary and pancreatic ductal epithelia and, in the embryonic period, in ureteric bud, biliary and pancreatic ducts and salivary gland. Immunofluorescence and immunoelectron microscopy studies suggest that polyductin localizes to primary apical cilium, apical membrane and cytoplasm of collecting duct cells. Our data indicate that PKHD1 codifies membrane and soluble isoforms

O gene PKHD1, mutado na doença renal policística autossômica recessiva, apresenta um padrão de splicing complexo associado a múltiplos transcritos alternativos. Neste trabalho estudamos o perfil de expressão de seu produto, poliductina. Análises por western blot revelaram produtos putativos de membrana de > 440 kDa e ~230 kDa, e de ~140 kDa em frações solúveis de rim, fígado e pâncreas. Estudos imunoistoquímicos mostraram marcação em ductos coletores renais e porção ascendente espessa da alça de Henle, em epitélios ductais biliar e pancreático e, no período embrionário, em broto ureteral, ductos biliar e pancreático e glândula salivar. Análises por imunofluorescência e microscopia imunoeletrônica sugerem que poliductina se localize em cílio apical primário, membrana apical e citoplasma de células do ducto coletor. Nossos resultados indicam que PKHD1 codifica isoformas de membrana e solúveis / PKHD1, the gene mutated in autosomal recessive polycystic kidney disease, presents a complex splicing pattern, associated with multiple alternative transcripts. In this work we have studied the expression profile of its product, polyductin. Western blot analysis revealed putative membrane products of > 440 kDa and 230 kDa, and of ~140 kDa in soluble fractions in kidney, liver and pancreas. Immunohistochemistry studies showed staining in renal collecting duct and thick ascending limb of Henle, in biliary and pancreatic ductal epithelia and, in the embryonic period, in ureteric bud, biliary and pancreatic ducts and salivary gland. Immunofluorescence and immunoelectron microscopy studies suggest that polyductin localizes to primary apical cilium, apical membrane and cytoplasm of collecting duct cells. Our data indicate that PKHD1 codifies membrane and soluble isoforms

Nous présentons ici la description clinique et génétique d’un syndrome neurocutané unique. Le laboratoire du Dr Cossette a entrepris la caractérisation clinique et génétique d'une famille canadienne-française qui a été identifiée par les Drs Giroux et Barbeau en 1972 et qui comprend plus de 100 personnes sur six générations. Les membres atteints de cette famille présentent des lésions typiques d'érythrokératodermie (EK) (OMIM 133190, EKV1 et EKV2), associées à une ataxie spinocérébelleuse pure. Dans cette famille, l'ataxie est caractérisée par des troubles de la coordination et de la démarche causés par une dégénérescence du cervelet et de la moelle épinière. Cette ataxie est transmise selon un mode autosomique dominant. Une étude antérieure de cette variante d'EK avec ataxie avait suggéré une liaison sur le chromosome 1p34-p35, soit la même région que les formes EKV de type 1 et 2, causées respectivement par des mutations dans les gènes connexin-31 (GJB3; OMIM 603324) et connexin-30.3 (GJB4; OMIM 605425). Cependant, aucune mutation n'a été retrouvée dans ces gènes pour la famille canadienne-française. Nous avons récemment recontacté la famille et effectué des examens détaillés, incluant une imagerie par résonance magnétique (IRM) et un électromyogramme (EMG). Les manifestations neurologiques des individus atteints sont compatibles avec une nouvelle forme d’ataxie cérébelleuse pure à transmission autosomique dominante (ADCA de type III dans la classification de Harding) que nous avons appelée SCA34. Une cartographie complète du génome nous a permis de localiser le gène SCA34 sur le chromosome 6p12.3-q16.2. Également, en collaboration avec les Drs Alexis Brice (Hôpital Pitié-La Salpêtrière, Paris) et Alfredo Brusco (Hôpital San Giovanni Battista di Torino, Italie), nous avons confirmé que trois autres familles européennes avec SCA inexpliquée étaient également liées au locus SCA34. Notre laboratoire a récemment entrepris la recherche des mutations responsables de SCA34. Les résultats de ce criblage de gènes candidats sont présentés dans le chapitre 3 de cette thèse. / We present here the clinical and genetic description of a unique neuro-cutaneous syndrome. Dr. Cossette’s laboratory began the clinical and genetic characterization of a French-Canadian family who was identified by Drs. Giroux and Barbeau in 1972 and includes more than 100 people over six generations. The affected members of this family have typical lesions of erythrokeratodermia (EK) (OMIM 133190, and EKV1 EKV2), associated with pure spinocerebellar ataxia. In this family, the clinical phenotype is characterized by gait ataxia caused by degeneration of the cerebellum and spinal cord and the pattern of inheritance is compatible with an autosomal dominant trait. In a previous study of this variant of ataxia with EK, putative linkage was found on chromosome 1p34-p35, the same chromosomal region of EKV1 and EKV2 that are respectively caused by mutations in the connexin-31 gene (GJB3, OMIM 603324) and connexin -30.3 (GJB4, OMIM 605425). However, no mutations have been found in these latter genes for the French-Canadian family. We recently contacted the family and carried out detailed examinations, including a magnetic resonance imaging (MRI) and electromyography (EMG). Neurological manifestations of affected individuals are consistent with a new form of pure autosomal dominant cerebellar ataxia, (ADCA type III in the classification of Harding) that we named SCA34. A whole genome scan allowed us to map the gene on chromosome 6p12.3-q16.2. Interestingly, in collaboration with Dr. Alexis Brice (Hôpital Pitié-La Salpêtrière, Paris), and Alfredo Brusco (San Giovanni Battista Hospital, Turin, Italy), we found that three additional European families with unexplained SCA were also linked to the SCA34 locus. Our laboratory has recently begun the search for mutations causing SCA34. The results of this screening of candidate genes are presented in Chapter 3 of this thesis.

Student Number : 9807768F - MSc (Med) research report - Faculty of Health Sciences / Fanconi anaemia (FA) is an autosomal recessive disorder characterised by aplastic anaemia (AA) and a high risk of developing acute myeloid leukaemia (AML). It is unknown whether heterozygote carriers are also predisposed to developing these disorders. The black South African population group is ideal for FA mutation screening because the presence of a founder mutation, FANCG 637-643, makes screening relatively straight forward. Three individuals with AML (115 screened) and one with AA (78 screened) were found to be heterozygous for the black South African founder mutation. From our data it seems unlikely that this mutation places heterozygous carriers of the mutation at high risk of developing AML or AA. Three children with AA out of 26 screened, were homozygous for the mutation. This finding reiterates the importance of screening all children with AA for FA. The frequency of certain congenital abnormalities in black South African FA patients was compared to patients described by other research groups. The frequencies of the abnormalities were similar to other FANCG cohorts described but significant differences to a group of FA patients from unspecified complementation groups were found. This difference could be because different complementation groups are associated more or less strongly with specific abnormalities. It was found previously that particular congenital abnormalities in FA patients are associated with a poor haematological outcome. We concluded that black South African FANCG patients have a high risk of early development of AA even though they do not have a high frequency of congenital abnormalities.

A doença renal policística autossômica dominante (DRPAD) é uma das doenças hereditárias humanas mais comuns, apresentando uma prevalência de 1:543 a 1:4000 em diferentes populações. Esta enfermidade é essencialmente causada por mutação em um de dois genes: PKD1 (Polycystic Kidney Disease 1) ou PKD2. A maior parte dos casos decorre de mutações em PKD1. O tabagismo foi associado independentemente com progressão da doença renal na DRPAD. Em um estudo recente, contudo, tabagismo não influenciou a sobrevida renal nesta doença. Vários estudos também revelam que a deficiência em PKD1/Pkd1 ou PKD2/Pkd2 se associam a disfunção cardíaca. Para analisar os efeitos potenciais da exposição crônica ao tabagismo sobre os fenótipos renal e cardíaco associados à DRPAD, utilizamos como modelo experimental um animal cístico ortólogo fenotipicamente semelhante à DRPAD humana, o camundongo Pkd1flox/flox:Nestincre. Animais machos foram expostos ao tabagismo da concepção até a idade de 18 semanas. Os níveis séricos de ureia foram mais elevados em camundongos Pkd1flox/flox:Nestincre fumantes (CIF) que em Pkd1flox/flox:Nestincre não fumantes (CI) e em animais Pkd1flox/flox fumantes (NCF) que em Pkd1flox/flox não fumantes (NC). O índice cístico renal foi maior nos animais CIF que CI. A taxa de proliferação de células de revestimento cístico, mas não de apoptose, mostrou-se aumentada em camundongos CIF comparados a CI. As taxas de proliferação e apoptose de células tubulares renais foram mais altas nos rins CI que NC e nos rins CIF que NCF. Observamos, ainda, tais taxas mais elevadas em NCF que NC. Camundongos CIF desenvolveram maior índice de fibrose renal que CI. O mesmo efeito do tabagismo foi detectado em camundongos NCF, porém sua intensidade foi muito menor. As expressões renais de Il1b e Tgfb1 não diferiram significantemente entre os grupos. Os níveis renais de glutationa foram mais baixos em camundongos CIF que CI e em animais NCF que NC. A fração de ejeção do ventrículo esquerdo foi reduzida em camundongos CIF e CI em comparação a NC, mas não foi detectada diferença entre CIF e CI. Parâmetros cardíacos estruturais também se mostraram alterados em animais CIF comparados a CI e NCF. Interessantemente, strain circunferencial e a taxa de strain circunferencial foram menores em camundongos CIF que CI e NCF. A pressão arterial média foi mais baixa em animais CIF que CI hipertensos, atingindo níveis que não diferiram de NC e NCF. A taxa de apoptose em tecido cardíaco foi maior em animais CI que NC, mas não diferiu entre CIF e CI nem entre NCF e NC. A taxa de fibrose cardíaca foi mais elevada em camundongos CIF e CI que NC e NCF. Os grupos não diferiram quanto à sobrevida em 18 semanas, mas o peso corporal foi menor em animais CIF que CI em 16 semanas. Nossos achados indicam que a exposição crônica ao tabagismo desde a concepção agravou os fenótipos renal e cardíaco de camundongos císticos deficientes em Pkd1. Dadas as similaridades entre a DRPAD humana e o modelo animal ortólogo estudado, os efeitos deletérios do tabagismo observados devem provavelmente se aplicar a pacientes com DRPAD / Autosomal dominant polycystic kidney disease (ADPKD) is one of the most common human inherited diseases, with a prevalence of 1:543 to 1:4000 in different populations. This disease is essentially caused by mutation in one of two genes: PKD1 (Polycystic Kidney Disease 1) or PKD2. Most cases result from mutations in PKD1. Smoking has been independently associated with renal disease progression in ADPKD. In a recent study, however, smoking did not influence renal survival in this disease. Several studies also revealed that PKD1/Pkd1 or PKD2/Pkd2 deficiency are associated with cardiac dysfunction. To analyze the potential effects of chronic exposure to smoking on renal and cardiac phenotypes associated with ADPKD, we used an orthologous cystic animal phenotypically similar to human ADPKD as experimental model, the Pkd1flox/flox:Nestincre mouse. Male animals were exposed to smoking from conception to the age of 18 weeks. Serum urea nitrogen was higher in Pkd1flox/flox:Nestincre submitted to smoking (CYS) than in Pkd1flox/flox:Nestincre not submitted to smoking (CY) and in Pkd1flox/flox submitted to smoking (NCS) than in Pkd1flox/flox not submitted to smoking (NC). Renal cystic index was higher in CYS animals than CY. Proliferation rate of cyst-lining cells, but not apoptotic rate, was increased in CYS mice compared to CY. The proliferation and apoptotic rates of renal tubular cells were higher in CY kidneys than NC and in CYS kidneys than NCS. We also observed increase in such rates in NCS compared to NC. CYS mice developed a higher renal fibrosis index than CY animals. Although the same effect of smoking was detected in NCS mice, its intensity was much lower than in cystic animals. Renal expression of Il1b e Tgfb1 did not significantly differ among the groups. Renal levels of glutathione were lower in CYS mice than CY and in NCS animals than NC. Left ventricular ejection fraction was reduced in CYS and CY mice compared to NC, but no difference was detected between CYS and CY. Cardiac structural parameters were also altered in CYS animals in comparison to CY and NCS. Interestingly, circumferential strain and circumferential strain rate were lower in the CYS mice than CY and NCS. MAP was lower in CYS animals than in the hypertensive CY mice, reaching normotensive levels that did not differ from NC and NCS. The apoptotic rate in cardiac tissue was higher in CY animals than NC, but did not differ between CYS and CY and between NCS and NC. The rate of cardiac fibrosis was also higher in CYS and CY mice than in NC and NCS. Survival at 18 weeks did not differ among the groups but body weight was lower in CYS animals than CY at 16 weeks of age. Our findings show that chronic exposure to smoking since conception aggravated the renal and cardiac phenotypes of Pkd1-deficient cystic mice. Given the similarities between human ADPKD and the studied orthologous animal model, the observed deleterious effects of smoking are likely to apply to ADPKD patients

Hemos analizado la variabilidad genética de una zona no codificante autosómica, el pseudogén homólogo al gen de la glucocerebrosidasa (psGBA). Parte del análisis se ha realizado desde la perspectiva de la genética de poblaciones humanas. Desde un punto de vista más genómico hemos establecido la dinámica de la región, a fin de entender las causas del espectro de variación. Hemos analizado el papel de la mutación, recombinación, conversión génica y, especialmente, selección. Por otra parte, psGBA es importante en la producción de alelos complejos GBA-psGBA, que provocan los tipos más severos de la enfermedad de Gaucher. Mostramos cómo el conocimiento de la variabilidad en psGBA ayuda al reconocimiento de estos alelos complejos. Finalmente, con los datos de variabilidad de dos regiones parálogas situadas en la misma región cromosómica (gen GBA / pseudogen psGBA) hemos comparado los patrones de mutación que presenta una misma secuencia bajo diferentes presiones selectivas. / We have analyzed the genetic variability in a non-coding autosomal region, the pseudogene homologous to the glucocerebrosidase gene (psGBA).Part of the analysis has been performed from the human populations point of view.From a more genomic perspective, we have established the region dynamics in order to understand the causes of the variability pattern. We have analyzed the role of mutation, recombination, gene conversion and, especially, selection.On the other hand, psGBA is important in the production of complex alleles GBA-psGBA, that lead to the most severe types of Gaucher disease. We show how the knowledge of psGBA variability helps to the identification of those complex alleles.Last, from the variability data from two paralogous regions located in the same chromosomal region (GBA gene /psGBA pseudogene) we have compared the mutation patterns shown by the same sequence under different selective pressures.

Nous présentons ici la description clinique et génétique d’un syndrome neurocutané unique. Le laboratoire du Dr Cossette a entrepris la caractérisation clinique et génétique d'une famille canadienne-française qui a été identifiée par les Drs Giroux et Barbeau en 1972 et qui comprend plus de 100 personnes sur six générations. Les membres atteints de cette famille présentent des lésions typiques d'érythrokératodermie (EK) (OMIM 133190, EKV1 et EKV2), associées à une ataxie spinocérébelleuse pure. Dans cette famille, l'ataxie est caractérisée par des troubles de la coordination et de la démarche causés par une dégénérescence du cervelet et de la moelle épinière. Cette ataxie est transmise selon un mode autosomique dominant. Une étude antérieure de cette variante d'EK avec ataxie avait suggéré une liaison sur le chromosome 1p34-p35, soit la même région que les formes EKV de type 1 et 2, causées respectivement par des mutations dans les gènes connexin-31 (GJB3; OMIM 603324) et connexin-30.3 (GJB4; OMIM 605425). Cependant, aucune mutation n'a été retrouvée dans ces gènes pour la famille canadienne-française. Nous avons récemment recontacté la famille et effectué des examens détaillés, incluant une imagerie par résonance magnétique (IRM) et un électromyogramme (EMG). Les manifestations neurologiques des individus atteints sont compatibles avec une nouvelle forme d’ataxie cérébelleuse pure à transmission autosomique dominante (ADCA de type III dans la classification de Harding) que nous avons appelée SCA34. Une cartographie complète du génome nous a permis de localiser le gène SCA34 sur le chromosome 6p12.3-q16.2. Également, en collaboration avec les Drs Alexis Brice (Hôpital Pitié-La Salpêtrière, Paris) et Alfredo Brusco (Hôpital San Giovanni Battista di Torino, Italie), nous avons confirmé que trois autres familles européennes avec SCA inexpliquée étaient également liées au locus SCA34. Notre laboratoire a récemment entrepris la recherche des mutations responsables de SCA34. Les résultats de ce criblage de gènes candidats sont présentés dans le chapitre 3 de cette thèse. / We present here the clinical and genetic description of a unique neuro-cutaneous syndrome. Dr. Cossette’s laboratory began the clinical and genetic characterization of a French-Canadian family who was identified by Drs. Giroux and Barbeau in 1972 and includes more than 100 people over six generations. The affected members of this family have typical lesions of erythrokeratodermia (EK) (OMIM 133190, and EKV1 EKV2), associated with pure spinocerebellar ataxia. In this family, the clinical phenotype is characterized by gait ataxia caused by degeneration of the cerebellum and spinal cord and the pattern of inheritance is compatible with an autosomal dominant trait. In a previous study of this variant of ataxia with EK, putative linkage was found on chromosome 1p34-p35, the same chromosomal region of EKV1 and EKV2 that are respectively caused by mutations in the connexin-31 gene (GJB3, OMIM 603324) and connexin -30.3 (GJB4, OMIM 605425). However, no mutations have been found in these latter genes for the French-Canadian family. We recently contacted the family and carried out detailed examinations, including a magnetic resonance imaging (MRI) and electromyography (EMG). Neurological manifestations of affected individuals are consistent with a new form of pure autosomal dominant cerebellar ataxia, (ADCA type III in the classification of Harding) that we named SCA34. A whole genome scan allowed us to map the gene on chromosome 6p12.3-q16.2. Interestingly, in collaboration with Dr. Alexis Brice (Hôpital Pitié-La Salpêtrière, Paris), and Alfredo Brusco (San Giovanni Battista Hospital, Turin, Italy), we found that three additional European families with unexplained SCA were also linked to the SCA34 locus. Our laboratory has recently begun the search for mutations causing SCA34. The results of this screening of candidate genes are presented in Chapter 3 of this thesis.

Indiana University-Purdue University Indianapolis (IUPUI) / In the past, the present and the future, Lebanon has been an important link between the East and the West. It was always known as the ‘Switzerland of the East’. Over the years, it was a hotspot for different civilizations that uniquely shaped the genomic backbone of the current Lebanese. It is also a good representation of genetically admixed individuals with diverse phenotype characteristics and unique features. Lebanon, quite like other Middle Eastern populations, lacks sufficient genetic studies that helps to better comprehend the complex genomic composition of different traits and diseases. The lack of good representation of the Middle East and North Africa (MENA) region in global studies has led to ambiguity in discovering special ancestry markers and patterns in the Lebanese genome. Yet, in this study, a thorough investigation into a Lebanese collection shows new patterns that potentially would be helpful in forensic and genealogical applications. The investigation into the autosomal and Y-STRs revealed unique alleles that would be valuable in future forensic investigation analysis. In addition, the assessment of phenotype prediction models to predict eye, hair and skin color showed promising results in terms of prediction performance. Those results encourage the future use of intelligence tools in the regions that in return would aid in serving justice and furthering science research. In fact, ancestry and genetic distance studies confirms the presence of admixture within Lebanon between Europe and North Africa. / 2029-06-01